IRR_Appendix_B_Quality_Specifications.pdf

CDC International Reagent Resource

Appendix B – Quality Specifications

Table of Contents

1. Live Influenza Viruses

2. Live Influenza Viruses (sequencing grade)

3. Beta-propiolactone (BPL) Inactivated Seasonal Human Influenza Viruses

4. Genomic RNA from Influenza Virus

5. Ferret Antisera to Seasonal Human Influenza Viruses

6. Ferret Antisera to Influenza Candidate Vaccine Viruses (CVV)

7. Influenza Recombinant Proteins

8. Cell Lines for Influenza Virus Propagation

9. Hybridomas Cell Line Cultivation

10. Monoclonal Antibody Purification

11. Influenza Reagent Kit for Identification of Influenza Isolates (“WHO Kit”)

12. Influenza Immunofluorescence Reagent Kit for Identification of Influenza Isolates

(“WHO IFA Kit”)

13. Influenza Real-time RT-PCR Kits (In Vitro Diagnostic)

14. Influenza Real-time RT-PCR Kits (Research Use Only)

15. Influenza Multi-segment Reverse Transcription PCR Kits

16. Influenza Real-time RT-PCR Sets

17. Influenza Real-time RT-PCR Controls

18. Influenza Reagent Panels

19. Bacterial Pathogen RT-PCR Detection Kits

20. Atypical Respiratory Pathogens RT-PCR Detection Kits

21. Streptococcus RT-PCR Typing Kit

22. Bacterial Pathogen RT-PCR Typing Kit

23. Streptococcus Conventional PCR Kit

24. Streptococcus RT-PCR Positive Controls

25. Bordetella RT-PCR Controls

26. Bacterial Pathogen RT-PCR Positive Controls, Simple Prep

27. Bacterial Pathogen Reference Strains

28. Cell Lines for Poliovirus Propagation

29. Norovirus Real-time RT-PCR Kits

30. Rotavirus Genotyping RT-PCR Kit

31. Measles and Rubella Real-time RT-PCR Kit

32. Poliovirus ITD RT-PCR Kits

33. Vaccine-derived Poliovirus (VDPV) RT-PCR Kits

34. Enterovirus Quadruplex RT-PCR Kits

35. Non-Influenza Respiratory Virus Real-time RT-PCR Panel

36. MERS-CoV Real-time RT-PCR Panel

37. Non-Influenza Respiratory Virus Real-time RT-PCR Kit

38. Respiratory Virus RT-PCR Positive Controls

39. Norovirus Conventional RT-PCR Kit

40. Rotavirus Conventional RT-PCR Kit

41. Measles Genotyping PCR Kit

42. Rubella Genotyping PCR Kit

43. Rubella Diagnostic PCR Kit

44. Poliovirus Sequencing Kit

45. Enterovirus Reagent Panels

1. Live Influenza Viruses

Format Infectious human seasonal or animal origin influenza virus in (a) clarified allantoic fluid from infected specific pathogen free embryonated chicken eggs or (b) clarified supernatant from infected tissue culture cells

Container 1.8 mL external thread screw cap cryovial

Volume 0.5 mL

Storage -60°C or colder

Shipment Frozen on dry ice

Documentation Product Specification for each product. Certificates of Analysis for each lot produced.

Labeling All vial, box, and package insert proofs inspected and approved by CDC before printing.

Quality Control Tests and Acceptance Criteria

CDC may perform independent QC testing of functional performance of all IRR-manufactured reagents at bulk, post-fill and final production stages to confirm that reagents meet all defined acceptance criteria. Acceptance of noncomplying results will be at the discretion of the CDC COR, with technical input from CDC subject matter experts.

Test Results

Appearance Solution is clear.

Type/subtype identification and purity by RT-PCR of RNA.

Minimum of five PCR targets representing at least a universal

Influenza A, H1N1pdm09, H3N2, Influenza B Victoria lineage, and Influenza B Yamagata lineage. Additional subtype specific targets may also be included.

Exclusive detection of expected influenza virus type/subtype within 20-28 CT range using sequences or primers/probes provided by CDC

Identification by hemagglutinin gene sequencing. CDC may require additional genes to confirm reassortants.

< 1% base changes as compared to results detected upon receipt or after first distribution lot if additional quantities are needed

Hemagglutination titer using 0.5% turkey red blood cells ≥ Titer of CDC seed stock virus provided

Titer by CEID50 assay in embryonated chicken eggs ≥ Titer of CDC seed stock virus if provided

Titer by TCID50 assay in MDCK cells or FFU/ml ≥ Titer of CDC seed stock virus if provided

Sterility (14-day incubation)

Soybean-casein digest medium, 22.5°C ± 2.5°C, aerobic

Fluid thioglycollate medium, 32.5°C ± 2.5°C, anaerobic

No growth

Mycoplasma contamination

No DNA detected by PCR of extracted test article nucleic acid.

2. Live Influenza Viruses (sequencing grade)

Format Infectious human seasonal virus in (a) clarified allantoic fluid from infected specific pathogen free embryonated chicken eggs or (b) clarified supernatant from infected tissue culture cells

Container 1.8 mL external thread screw cap cryovial

Volume 0.5 mL

Storage -60°C or colder

Shipment Frozen on dry ice

Documentation Product Specification for each product. Certificates of Analysis for each lot produced.

Labeling All vial, box, and package insert proofs inspected and approved by CDC before printing.

Quality Control Tests and Acceptance Criteria

CDC may perform independent QC testing of functional performance of all IRR-manufactured reagents at bulk, post-fill and final production stages to confirm that reagents meet all defined acceptance criteria. Acceptance of noncomplying results will be at the discretion of the CDC COR, with technical input from CDC subject matter experts.

Test Results

Appearance Solution is clear.

Genomic amplification and next generation sequencing.

Presence of too many DI particles hamper the ability to effectively sequence the influenza genome.

Even coverage across each gene segment with depth of 1500x to 4000x. Uneven coverage represents the presence of DI particles.

Type/subtype identification and purity by RT-PCR of RNA.

Minimum of five PCR targets representing universal Influenza

A, H1N1pdm09, H3N2, Influenza B Victoria lineage, and

Influenza B Yamagata lineage. Additional subtype specific targets may also be included.

Exclusive detection of expected influenza virus type/subtype within 20-28 CT range using sequences or primers/probes provided by CDC

Identification by hemagglutinin gene sequencing. CDC may require additional genes to confirm reassortants.

< 1% base changes as compared to results detected upon receipt or after first distribution lot if additional quantities are needed

Hemagglutination titer using 0.5% turkey red blood cells ≥ Titer of CDC seed stock virus provided

Titer by CEID50 assay in embryonated chicken eggs ≥ Titer of CDC seed stock virus if provided

Titer by TCID50 assay in MDCK cells or FFU/ml ≥ Titer of CDC seed stock virus if provided

Sterility (14-day incubation)

Soybean-casein digest medium, 22.5°C ± 2.5°C, aerobic

Fluid thioglycollate medium, 32.5°C ± 2.5°C, anaerobic

No DNA detected by PCR of extracted test article nucleic acid.

3. Beta-propiolactone (BPL) Inactivated Seasonal Human Influenza Viruses

Format BPL-inactivated virus in either allatonic fluid or tissue culture supernatant, containing 0.1% sodium azide

Container 1.8 mL external thread screw cap cryovial

Volume 0.5 mL

Storage -60°C or colder

Shipment Frozen on dry ice

Documentation Product Specification for each product. Certificates of Analysis for each lot produced.

Labeling All vial, box, and package insert proofs inspected and approved by CDC before printing.

Quality Control Tests and Acceptance Criteria

CDC may perform independent QC testing of functional performance of all IRR-manufactured reagents at bulk, post-fill and final production stages to confirm that reagents meet all defined acceptance criteria. Acceptance of noncomplying results will be at the discretion of the CDC COR, with technical input from CDC subject matter experts.

Test Results

Appearance Solution is clear.

Type/subtype identification and purity by RT-PCR of RNA.

Minimum of five PCR targets representing universal Influenza

A, H1N1pdm09, H3N2, Influenza B Victoria lineage, and

Influenza B Yamagata lineage. Additional subtype specific targets may also be included.

Exclusive detection of expected influenza virus type/subtype within 45 PCR cycles using sequences or primers/probes provided by CDC

Reactivity by RT-PCR of RNA.

Minimum of five PCR targets representing universal Influenza

A, H1N1pdm09, H3N2, Influenza B Victoria lineage, and

Influenza B Yamagata lineage. Additional subtype specific targets may also be included.

≤ four Ct difference within 45 PCR cycles compared to original infectious starting material using sequences or primers/probes provided by

CDC

Hemagglutination titer using 0.5% turkey red blood cells ≥ Titer of CDC seed stock virus provided

Innocuity for egg passaged viruses

Undetectable virus growth in eggs with no positive HA result. A minimum of two consecutive passages is required.

Innocuity for cell passaged viruses

Undetectable virus growth in a cell culture assay with no positive infectious virus result. A minimum of three replicates is required.

Sterility (14-day incubation)

Soybean-casein digest medium, 22.5°C ± 2.5°C, aerobic

Fluid thioglycollate medium, 32.5°C ± 2.5°C, anaerobic

4. Genomic RNA from Influenza Virus

Format Genomic RNA extracted from a preparation of clarified allantonic fluid from specific pathogen free embryonated chicken eggs infected with influenza A or B virus

Container 0.5mL DNase-free, RNase-free cryovial

Volume 100 uL

Storage -60°C or colder

Shipment Frozen on dry ice

Documentation Product Specification for each product. Certificates of Analysis for each lot produced.

Labeling All vial, box, and package insert proofs inspected and approved by CDC before printing.

Quality Control Tests and Acceptance Criteria

CDC may perform independent QC testing of functional performance of all IRR-manufactured reagents at bulk, post-fill and final production stages to confirm that reagents meet all defined acceptance criteria. Acceptance of noncomplying results will be at the discretion of the CDC COR, with technical input from CDC subject matter experts.

Test Results

Appearance Solution is clear.

Concentration ≥ 5ng/uL

Type/subtype identification and purity by RT-PCR of RNA.

Minimum of five PCR targets representing universal Influenza

A, H1N1pdm09, H3N2, Influenza B Victoria lineage, and

Influenza B Yamagata lineage. Additional subtype specific targets may also be included.

Exclusive detection of expected influenza virus type/subtype within 45 PCR cycles using sequences or primers/probes provided by CDC

Undetectable virus growth in eggs with no positive HA result. A minimum of two consecutive passages is required.

with no positive infectious virus result. A minimum of three replicates is required.

OD260/OD280 Ratio 1.9 ± 0.1

5. Ferret Antisera to Seasonal Human Influenza Viruses

Format Heat-inactivated ferret antisera against live or inactivated influenza virus. Prior to infection, ferrets must be seronegative for antibodies against current seasonal H1N1 and H3N2 influenza A antigens and seasonal influenza B antigens. Sera will not be treated with receptor destroying enzyme (RDE).

Container 1.2 mL external thread screw cap cryovial

Volume 1.0 mL

Storage -20°C or colder

Shipment Frozen on dry ice

Documentation Product Specification for each product. Certificates of Analysis for each lot produced.

Labeling All vial, box, and package insert proofs inspected and approved by CDC before printing.

Quality Control Tests and Acceptance Criteria

CDC may perform independent QC testing of functional performance of all IRR-manufactured reagents at bulk, post-fill and final production stages to confirm that reagents meet all defined acceptance criteria. Acceptance of noncomplying results will be at the discretion of the CDC COR, with technical input from CDC subject matter experts.

Test Results

Appearance Solution is clear.

Sensitivity - Hemagglutination inhibition titers with influenza virus using 0.5% turkey red blood cells and

RDE treated serum from pre and post vaccinated ferrets

Pre-vaccinated ferret serum ≤ 40 to homologous virus and seasonal influenza virus strains

Post-vaccinated ferret serum ≥ 320 to homologous virus

Specificity - Hemagglutination inhibition assay with at least four antigens using 0.5% turkey red blood cells. Current homologous and heterologous antigens will be identified and/or provided by CDC.

≤ 40 to heterologous antigens

≥ 320 to homologous antigens

≥ 4-fold difference in HI titer between current homologous and heterologous antigens

Innocuity for serum generated from egg passaged viruses

Undetectable virus growth in eggs with no positive HA result. A minimum of two consecutive passages is required.

Innocuity for serum generated from cell passaged viruses

Undetectable virus growth in a cell culture assay with no positive infectious virus result. A minimum of three replicates is required.

6. Ferret Antisera to Influenza Candidate Vaccine Viruses (CVV)

Format Heat-inactivated ferret antisera against live or inactivated influenza virus. Prior to infection, ferrets must be seronegative for antibodies against current seasonal H1N1 and H3N2 influenza A antigens and seasonal influenza B antigens. Sera will not be treated with receptor destroying enzyme (RDE).

Container 1.2 mL external thread screw cap cryovial

Volume 1.0 mL

Storage -60°C or colder

Shipment Frozen on dry ice

Documentation Product Specification for each product. Certificates of Analysis for each lot produced.

Labeling All vial, box, and package insert proofs inspected and approved by CDC before printing.

Quality Control Tests and Acceptance Criteria

CDC may perform independent QC testing of functional performance of all IRR-manufactured reagents at bulk, post-fill and final production stages to confirm that reagents meet all defined acceptance criteria. Acceptance of noncomplying results will be at the discretion of the CDC COR, with technical input from CDC subject matter experts.

Test Results

Appearance Solution is clear.

Sensitivity - Hemagglutination inhibition titers with influenza virus using 0.5% turkey red blood cells and

RDE treated serum from pre and post infected ferrets

Pre-infected ferret serum ≤ 40 to homologous virus and seasonal influenza virus strains

Post-infected ferret serum ≥ 160 to homologous virus

Specificity - Hemagglutination inhibition assay with at least four antigens using 0.5% turkey red blood cells. Current homologous and heterologous antigens will be identified and/or provided by CDC.

≤ 40 to heterologous antigens

≥ 160 to homologous antigens

≥ 4-fold difference in HI titer between current homologous and heterologous antigens

Innocuity for serum generated from egg passaged viruses

Undetectable virus growth in eggs with no positive HA result. A minimum of two consecutive passages is required.

Innocuity for serum generated from cell passaged viruses

Undetectable virus growth in a cell culture assay with no positive infectious virus result. A minimum of three replicates is required.

Presence of viral nucleic acids by RT-PCR of RNA.

Minimum of five PCR targets representing at universal Influenza A, Influenza B Victoria lineage, and Yamagata lineages and high path subtypes (e.g.

H5, H7). Additional subtype specific targets may also be included.

No detection of influenza virus type/subtype within 45 PCR cycles using sequences or primers/probes provided by CDC

7. Influenza Recombinant Proteins

Format Recombinant external or internal influenza viral proteins from a CDC-provided baculovirus construct or a contractor/subcontractor generated construct, with or without a six-histidine tag

Container 1.2 mL external thread screw cap cryovial

Volume At least 250ug in 0.25mL

Storage -80°C ± 10ºC

Shipment Frozen on dry ice

Documentation Product Specification for each product. Certificates of Analysis for each lot produced.

Labeling All vial, box, and package insert proofs inspected and approved by CDC before printing.

Expression and purification strategies must be approved by CDC before production begins to ensure optimal protein folding.

Quality Control Tests and Acceptance Criteria

CDC may perform independent QC testing of functional performance of all IRR-manufactured reagents at bulk, post-fill and final production stages to confirm that reagents meet all defined acceptance criteria. Acceptance of noncomplying results will be at the discretion of the CDC COR, with technical input from CDC subject matter experts.

Test Results

Appearance Solution is clear.

SDS-PAGE (Coomassie Blue Densitometry)

Specific protein bands (e.g. HA1, HA2, HA0) account for ≥ 90% of total density

Identification confirmation by western blot, ELISA, or cleavage assay

Reactive with a protein specific antibody and/or a histidine tag

Gel filtration chromatography ≥ 80% tetramer

Concentration by Bradford Assay 1.00 mg per mL ± 0.25 mg per mL

8. Cell Lines for Influenza Virus Propagation

Format Viable cell culture suspension in cell growth media (90%) and DMSO (10%) cryopreservative

Container 1.8 mL external thread screw cap cryovial

Volume 1.0 mL

Storage -130°C or colder, in the vapor phase of a liquid nitrogen freezer

Shipment Frozen on dry ice

Documentation Product Specification for each product. Certificates of Analysis for each lot produced.

Labeling All vial, box, and package insert proofs inspected and approved by CDC before printing.

Quality Control Tests and Acceptance Criteria

CDC may perform independent QC testing of functional performance of all IRR-manufactured reagents at bulk, post-fill and final production stages to confirm that reagents meet all defined acceptance criteria. Acceptance of noncomplying results will be at the discretion of the CDC COR, with technical input from CDC subject matter experts.

Madin-Darby Canine Kidney (MDCK) Cells

Madin-Darby Canine Kidney (MDCK) SIAT 1 Cells

Madin-Darby Canine Kidney (MDCK) London Luc 9.1 Cells

Test Results

Growth Properties Adherent

Morphology Epithelial-like

Total Cell Count ≥ 1x106 cells per vial

Post-freeze viability ≥ 75%

Species identification by multiplex PCR amplification of cytochrome C oxidase subunit I gene

Species confirmed.

Identification by direct fluorescence microscopy (for cells transfected with an influenza A-inducible Renilla luciferase-based reporter only)

Green fluorescence observed.

Induction by luciferase activity by influenza A infection (for cells transfected with an influenza A-inducible Renilla luciferase-based reporter only)

Luciferase activity detected.

Sterility (14-day incubation)

Soybean-casein digest medium, 22.5°C ± 2.5°C, aerobic

Fluid thioglycollate medium, 32.5°C ± 2.5°C, anaerobic

9. Hybridomas Cell Line Cultivation

Format Hybrid cell lines produced by fusion of antibody-producing lymphocytes with tumor cells in order to generate highly specific monoclonal antibodies

Container 1.8 mL external thread screw cap cryovial

Volume 1.0 mL

Storage -130°C or colder, in the vapor phase of a liquid nitrogen freezer

Shipment Frozen on dry ice

Documentation Product Specification for each product. Certificates of Analysis for each lot produced.

Labeling All vial, box, and package insert proofs inspected and approved by CDC before printing.

Quality Control Tests and Acceptance Criteria

CDC may perform independent QC testing of functional performance of all IRR-manufactured reagents at bulk, post-fill and final production stages to confirm that reagents meet all defined acceptance criteria. Acceptance of noncomplying results will be at the discretion of the CDC COR, with technical input from CDC subject matter experts.

CDC will provide hybridoma for expansion.

Test Results

Morphology Lymphoid

Total Cell Count ≥ 2x106 cells per vial

Post-freeze viability ≥ 25% at initial thaw

≥ 75% at Day 3 in culture

Antibody Class Confirmed IgG or IgA only

Report IgG subtype

Sterility (14-day incubation)

Soybean-casein digest medium, 22.5°C ± 2.5°C, aerobic

Fluid thioglycollate medium, 32.5°C ± 2.5°C, anaerobic

No growth

Mycoplasma contamination No DNA detected by PCR of extracted test article

10. Monoclonal Antibody Purification

Format Monoclonal antibody specific to influenza virus, may be in solution or lyophilized

Container 1.2 mL external thread screw cap cryovial

Volume 0.25mg of purified monoclonal antibody in 0.5 mL

Storage -20°C or colder

Shipment Frozen on dry ice

Documentation Product Specification for each product. Certificates of Analysis for each lot produced.

Labeling All vial, box, and package insert proofs inspected and approved by CDC before printing.

Quality Control Tests and Acceptance Criteria

CDC may perform independent QC testing of functional performance of all IRR-manufactured reagents at bulk, post-fill and final production stages to confirm that reagents meet all defined acceptance criteria. Acceptance of noncomplying results will be at the discretion of the CDC COR, with technical input from CDC subject matter experts.

Test Results

Appearance Solution is clear.

Lyophilized cake is white or colorless.

Antibody Class Confirmed IgG or IgA only

Report IgG subtype

SDS-PAGE (Coomassie Blue Densitometry) Correct molecular weight for heavy and light chains

> 90% pure

Concentration by spectrophotometer at OD280 0.5 mg per mL ± 10%

Functional activity against inactivated viruses or antigens.

Current homologous and heterologous inactivated viruses or antigens will be identified and/or provided by CDC.

Reactive to antigen by western blot and/or ELISA.

Report reactivity dilution or concentration results.

ForteBio® Octect® binding analysis Report results.

Biacore™ analysis (binding affinity) Report results.

Filtration 0.22 um filter-sterilized

Sterility (14-day incubation)

Soybean-casein digest medium, 22.5°C ± 2.5°C, aerobic

Fluid thioglycollate medium, 32.5°C ± 2.5°C, anaerobic

No growth

Mycoplasma contamination No DNA detected by PCR of extracted test article

11. Influenza Reagent Kit for Identification of Influenza Isolates (“WHO

Kit”)

Format Assembled kit containing reagents for hemagglutination inhibition (HAI) testing of field isolates and serologic diagnosis of infection:

(1) BPL-inactivated influenza A or B antigens

(2) Ether-extracted influenza B antigens

(3) Influenza reference antisera

(4) Influenza normal control sera

(5) Receptor destroying enzyme II (RDEII), Influenza Virus from Accurate Chemical, catalog # YCC340122.

Kit includes one package insert

Container Custom box with foam insert

Storage 2 to 8°C

Shipment Room temperature

Documentation Product Specification for each product. Certificates of Analysis for each lot produced.

Labeling All vial, box, and package insert proofs inspected and approved by CDC before printing.

Quality Control Tests and Acceptance Criteria

CDC may perform independent QC testing of functional performance of all IRR-manufactured reagents at bulk, post-fill and final production stages to confirm that reagents meet all defined acceptance criteria. Acceptance of noncomplying results will be at the discretion of the CDC COR, with technical input from CDC subject matter experts.

BPL-inactivated influenza A or B antigens 10 mL in glass vial with crimp seal

Ether-extracted Influenza B antigens 5 mL in glass vial with crimp seal

Test Results

Type/subtype identification and purity by RT-PCR of RNA.

Minimum of five PCR targets representing universal

Influenza A, H1N1pdm09, H3N2, Influenza B Victoria lineage, and Influenza B Yamagata lineage. Additional subtype specific targets may also be included.

Exclusive detection of expected influenza virus type/subtype within 45 PCR cycles using sequences or primers/probes provided by CDC

Hemagglutination titer using 0.5% turkey red blood cells ≥ 1:128 for Influenza A Antigens

≥ 1:64 for Influenza B Antigens

Undetectable virus growth in eggs with no positive

HA result. A minimum of two consecutive passages is required.

with no positive infectious virus result. A minimum of three replicates is required.

Sterility (14-day incubation)

Soybean-casein digest medium, 22.5°C ± 2.5°C, aerobic

Fluid thioglycollate medium, 32.5°C ± 2.5°C, anaerobic

No growth

Bromelain-treated seasonal human influenza viruses for production of antisera

Minimum of 100 ug for each prime injection

Minimum of 30 ug for each boost injection

Test Results

Purity by Gel Electrophoresis Confirmation of HA cleavage and purity (absence of other influenza proteins, ie. NP, M1, etc.)

Concentration by Bradford Assay or equivalent 1.00 mg per mL ± 0.25 mg per mL

Hemagglutination titer using 0.5% turkey red blood cells Provide titer

CDC will use bromelain-treated viruses as prime and/or boost injections for production of antisera in goat, sheep or ferrets. Animals may be boosted up to three times. CDC will provide plasmapheresed antisera to the contractor as a bulk, to be aliquoted at 5mL/vial and lyophilized at the contractor’s facility.

Pre-vaccinated animal ≤ 40 to homologous antigen and seasonal influenza strains

Post-vaccinated animal serum ≥ 640 to homologous BPL-treated Influenza A WHO Kit antigens

Post-vaccinated animal serum ≥ 320 to homologous BPL-treated Influenza B WHO Kit antigens

≥ 4-fold difference in HI titer between current homologous and heterologous antigens

12. Influenza Immunofluorescence Reagent Kit for Identification of

Influenza Isolates (“WHO IFA Kit”)

Format Virtual panel containing four monoclonal antibodies for identification of influenza isolates using immunofluorescence staining

Container Virtual Panel

Storage 2 - 8°C

Shipment Room temperature

Documentation Product Specification for each product. Certificates of Analysis for each lot produced.

Labeling All vial, box, and package insert proofs inspected and approved by CDC before printing.

Quality Control Tests and Acceptance Criteria

CDC may perform independent QC testing of functional performance of all IRR-manufactured reagents at bulk, post-fill and final production stages to confirm that reagents meet all defined acceptance criteria. Acceptance of noncomplying results will be at the discretion of the CDC COR, with technical input from CDC subject matter experts.

Monoclonal Antibodies (mAbs) lyophilized Influenza A mAb (crossreactive pool) lyophilized Influenza B mAb (crossreactive pool) lyophilized Influenza A mAbs (pdmH1N1) lyophilized Influenza A mAbs (H3)

Volume

0.5 mL in glass vial with crimp seal

0.5 mL in glass vial with crimp seal

0.5 mL in glass vial with crimp seal

0.5 mL in glass vial with crimp seal

CDC will provide hybridoma cells for production of the Influenza A and B mAb crossreactive pools. The H1N1 and

H3 seasonal influenza mAbs may be purchased in bulk and then vialed and lyophilized at the contractor’s facility.

Test Results

Immunofluoresecence assay (IFA)

100% positivity at a ≥ 100-fold dilution by IFA against homologous antigen

< 1% positivity at a ≥ 300-fold dilution by IFA against heterologous influenza viruses

13. Influenza Real-time RT-PCR Kits (In Vitro Diagnostic)

Format Kits containing oligonucleotide primers and probes for the detection of RNA from contemporary human influenza A and B viruses using real-time RT-PCR technology:

(1) 3-6 primer/probe sets each set includes a forward primer, reverse primer, and probe

(2) positive control

(3) nucleic acid extraction control

Kit includes one package insert

Container Outer packaging – Corrugated box, Dimensions: 8.88” x 5.94” x 1.75”, Jarrett Industries

(Owings, MD), containing:

(1) Primers and Probes Box

(2) Controls Box (positive control, extraction controls)

Inner packaging – Corrugated box, 8.25” x 2.25” x 1.25”, Jarrett Industries (Owings, MD), containing:

Vials – 2.0 mL Sarstedt with O-ring, sterile, cat #: 72-694-006

Storage -20°C or colder

Shipment Frozen on dry ice

Documentation All batch record documentation related to labeling, kit assembly, inspection, and storage shall be provided to CDC upon completion of task.

Labeling Box Label: ULINE, cat # S-6784, Dimensions: 3.00”W x 2.00”H, Synthetic Transfer

Printable Glossy White Polyester. Ribbon: ULINE, 4.02” x 1345”, Thermal Transfer Wax or

Wax/Resin (Black Resin)

Vial Label: Anthony-Lee Associates, Inc. Freezer Bondz, cat # CTL-168-492-AD, Dimensions: 0.75” x 1.68”. Thermal Transfer Printable Glossy White Polyester Label

Stock. Ribbon: Partnered Print Solutions Catalog Number: FST051F1, 2.01” x 13.45”, Thermal Transfer, Wax or Wax/Resin (Black FH Wax)

CDC will provide all inner and outer box label templates.

All vial, box, and package insert proofs inspected and approved by CDC before printing.

Proofs must be provided as hard copies (not electronic).

The CDC Influenza Division’s Human Influenza Virus Real-time RT-PCR Diagnostic panels (IVD) are diagnostic assays subject to regulatory requirements established by the US Food and Drug Administration (FDA). Current good manufacturing practices (CGMPs) include critical requirements related to the methods, facilities, and controls used for manufacture as well as packaging, labeling, storage and distribution of medical devices.

The Influenza Division will independently coordinate manufacture of RT-PCR primer/probe sets, positive controls, and extractions controls. Reagents will be transferred to IRR for labeling, final kit assembly, storage, and distribution only.

Influenza A/B Typing

3 primer and probe sets

17 extraction control vials

1 positive control vial

Influenza A Subtyping 6 primer and probe sets

1 positive control vial

Influenza B Genotyping 3 primer and probe sets

1 positive control vial

Avian Influenza Subtyping

3 primer and probe sets

17 extraction control vials

1 positive control vial

Quality Control Tests and Acceptance Criteria

CDC may perform independent QC testing of functional performance of all IRR-manufactured reagents at bulk, post-fill and final production stages to confirm that reagents meet all defined acceptance criteria. Acceptance of noncomplying results will be at the discretion of the CDC COR, with technical input from CDC subject matter experts.

Test Results

Label Printing 100% label check with 100% acceptance, including label placement on vial

If contractor demonstrates compliance in 10 acceptable lots with no defects, sampling may be reduced in accordance with ANSI/ASQ Z1.4-2008, Sampling Procedures and Tables for Inspection by Attributes.

Package Insert Printing Inserts must be printed in color, double-sided, and spiral bound and fit inside final packaging.

100% label check with 100% acceptance, including a smear test

Final Kit Assembly Quality audit, in accordance with ANSI/ASQ Z1.4-2008, Sampling

Procedures and Tables for Inspection by Attributes using a general inspection level II, AQL .065 must be performed and accepted by Quality

Assurance before samples are sent to CDC for inspection. Inspection shall include a label check and placement of vials in the appropriate inner boxes.

Maintain Storage Cold Chain 100% acceptance of functional performance of QC samples after assembly.

Ability of shipping packaging to maintain reagents frozen must be validated prior to initial use, and on an annual basis thereafter.

Non-confirming events (NCE) Requires immediate notification of CDC. All NCE documentation must be provided to CDC including investigation records, supporting data, and corrective actions. Resumption of activities dependent upon CDC approval/release. Additional samples may be requested for QC testing before resumption of distribution activities.

14. Influenza Real-time RT-PCR Kits (Research Use Only)

Format Kits containing oligonucleotide primers and probes for the detection of RNA from contemporary human influenza A and B viruses using real-time RT-PCR technology:

(1) 3-6 primer/probe sets

Each set includes a forward primer, reverse primer, and probe.

Kit includes one package insert

Container Outer packaging – Corrugated Box, Dimensions: 8.25” x 2.25” x 1.25”, Jarrett Industries

(Owings, MD), containing:

(1) Primers and Probes Box

Vials – 2.0 mL Sarstedt with O-ring, sterile, cat #: 72-694-006

Storage -20°C or colder

Shipment Frozen on dry ice

Documentation All batch record documentation related to labeling, kit assembly, inspection, and storage shall be provided to CDC upon completion of each lot.

Certificates of Analysis for each lot will be provided by CDC.

Labeling Box Label: ULINE, cat # S-6784, Dimensions: 3.00”W x 2.00”H, Synthetic Transfer

Printable Glossy White Polyester. Ribbon: ULINE, 4.02” x 1345”, Thermal Transfer Wax or

Wax/Resin (Black Resin)

Vial Label: Anthony-Lee Associates, Inc. Freezer Bondz, cat # CTL-168-492-AD, Dimensions: 0.75” x 1.68”. Thermal Transfer Printable Glossy White Polyester Label

Stock. Ribbon: Partnered Print Solutions Catalog Number: FST051F1, 2.01” x 13.45”, Thermal Transfer, Wax or Wax/Resin (Black FH Wax)

CDC will provide all inner and outer box label templates.

All vial, box, and package insert proofs inspected and approved by CDC before printing.

Proofs must be provided as hard copies (not electronic).

The CDC Influenza Division’s Human Influenza Virus Real-time RT-PCR Diagnostic panels (RUO) must be complaint with current good manufacturing practices (CGMPs), including critical requirements related to the methods, facilities, and controls used for manufacture as well as packaging, labeling, storage and distribution.

The Influenza Division will independently coordinate manufacture of RT-PCR primer/probe sets, positive controls, and extractions controls. Reagents will be transferred to IRR for labeling, final kit assembly, storage, and distribution only.

Influenza A/B Typing 3 primer and probe sets

Influenza A Subtyping 6 primer and probe sets

Influenza B Genotyping 3 primer and probe sets

Avian Influenza Subtyping 3 primer and probe sets

Quality Control Tests and Acceptance Criteria

CDC may perform independent QC testing of functional performance of all IRR-manufactured reagents at bulk, post-fill and final production stages to confirm that reagents meet all defined acceptance criteria. Acceptance of noncomplying results will be at the discretion of the CDC COR, with technical input from CDC subject matter experts.

Test Results

Label Printing 100% label check with 100% acceptance, including label placement on vial

If contractor demonstrates compliance in 10 acceptable lots with no defects, sampling may be reduced in accordance with ANSI/ASQ Z1.4-2008, Sampling

Procedures and Tables for Inspection by Attributes.

Package Insert Printing 100% label check with 100% acceptance, including a smear test

Final Kit Assembly Quality audit, in accordance with ANSI/ASQ Z1.4-2008, Sampling Procedures and Tables for Inspection by Attributes using a general inspection level II, AQL .065 must be performed and accepted by Quality Assurance before samples are sent to CDC for inspection. Inspection shall include a label check and placement of vials in the appropriate inner boxes.

Maintain Storage Cold Chain 100% acceptance of functional performance of QC samples after assembly.

Ability of shipping packaging to maintain reagents frozen must be validated prior to initial use, and on an annual basis thereafter.

Non-confirming events (NCE) Requires immediate notification of CDC. All NCE documentation must be provided to CDC including investigation records, supporting data, and corrective actions. Resumption of activities dependent upon CDC approval/release. Additional samples may be requested for QC testing before resumption of distribution activities.

15. Influenza Multi-segment Reverse Transcription PCR Kits

Format Kits containing lyophilized oligonucleotide primer cocktails for the detection of RNA from contemporary human influenza A and B viruses using multiplex reverse transcription (MRT) technology:

(1) One 3 primer pool

(2) One 13 primer pool

Kit includes one package insert

Container Outer packaging – Corrugated Box, Dimensions: 8.25” x 2.25” x 1.25”, Jarrett Industries

(Owings, MD), containing:

(1) Primer Box

Vials – 2.0 mL Sarstedt with O-ring, sterile, cat #: 72-694-006

Storage 2 to 8°C

Shipment 2 to 8°C

Documentation Certificates of Analysis for each lot produced

Labeling All vial, box, and package insert proofs inspected and approved by CDC before printing.

CDC will provide primers/probes for comparing the functional performance of the first lot manufactured by IRR in order to establish initial assay performance.

Primers: 20-30 nucleotides Purification

(primers)

Quantity per Vial

(forward, reverse)

Influenza Salt-Free 15 nmol, 15 nmol

* Preliminary configurations only, final production values may be adjusted after final assay optimization.

Quality Control Tests and Acceptance Criteria

CDC may perform independent QC testing of functional performance of all IRR-manufactured reagents at bulk, post-fill and final production stages to confirm that reagents meet all defined acceptance criteria. Acceptance of noncomplying results will be at the discretion of the CDC COR, with technical input from CDC subject matter experts.

Test Results

Sequence identity Verification of correct oligonucleotide sequence, including presence of all applicable sequence degeneracies

Sequence modifications Presence of correct modifications as applicable

Oligonucleotide purity ≥ 90% as determined by RNase Free HPLC purification

Molecular weight ≤ 2.0% of theoretical molecular weight of the product including all applicable sequence degeneracies

Functional performance Specific detection of target pathogen

Ct results within range specified by CDC

16. Influenza Real-time RT-PCR Sets

Format A la carte sets containing oligonucleotide primers and probes for the qualitative detection and identification of influenza viruses using RT-PCR technology:

(1) One forward primer, one reverse primer and one probe per set

Each set includes one package insert.

Container Fisher Nunc Transport Boxes for Cryoline, Fisher Scientific Cat # 12-565-184, packaged inside clear resealable bag

Contractor may propose alternative packaging options for approval by CDC as needed.

Storage -20°C or colder

Shipment Frozen on dry ice

Documentation Certificates of Analysis for each lot produced

Labeling All vial, box, and package insert proofs inspected and approved by CDC before printing.

CDC will provide primers/probes for comparing the functional performance of the first lot manufactured by IRR in order to establish initial assay performance.

Primers: 20-28 nucleotides

Probes: 35-45 nucleotides 5' mod internal mod 3' mod

Purification

(primers/probes)

Quantity per Vial

(forward, reverse, probe)

Influenza BHQ-1

BHQ-1

or Zen

BHQ-1 or

Iowa Black RP HPLC/Dual HPLC 20 nmol, 20nmol, 5 nmol

* Preliminary configurations only, final production values may be adjusted after final assay optimization.

Quality Control Tests and Acceptance Criteria

CDC may perform independent QC testing of functional performance of all IRR-manufactured reagents at bulk, post-fill and final production stages to confirm that reagents meet all defined acceptance criteria. Acceptance of noncomplying results will be at the discretion of the CDC COR, with technical input from CDC subject matter experts.

Test Results

Sequence identity Verification of correct oligonucleotide sequence, including presence of all applicable sequence degeneracies

Sequence modifications Presence of correct fluorophore, quencher, and other modifications as applicable.

Oligonucleotide purity ≥ 90% as determined by analytical HPLC or equivalent

Molecular weight ≤ 2.0% of theoretical molecular weight of the product including all applicable sequence degeneracies

Functional performance Specific detection of target and no cross reactivity to non-target sequences or viruses of interest using a panel of virus isolates provided by

CDC. Ct results within range specified by CDC.

17. Influenza Real-time RT-PCR Controls

Format Noninfectious (beta-propiolactone inactivated) positive control materials

(influenza virus preparation) and/or cultured human cells (A549) suspended in

0.01 M phosphate buffered saline (PBS) at pH 7.2-7.4.

Container 2.0 mL Sarstedt Packing Vial with O-ring, cat # 72-694-006

Storage -20°C or colder for liquid or rehydrated controls

2 to 8°C for lyophilized controls

Shipment Frozen on dry ice for liquid controls

Room temperature for lyophilized controls

Documentation Product Specification for each panel. Certificates of Analysis for each component lot produced.

Labeling All vial, box, and package insert proofs inspected and approved by CDC before printing.

The CDC Influenza Division’s real-time RT-PCR positive controls and extraction controls must be complaint with current good manufacturing practices (CGMPs), including critical requirements related to the methods, facilities, and controls used for manufacture as well as packaging, labeling, storage and distribution.

The Influenza Division will independently coordinate manufacture of positive controls, including lyophilization if applicable. Reagents will be transferred to IRR for labeling, final kit assembly, storage, and distribution only.

Quality Control Tests and Acceptance Criteria

CDC may perform independent verification QC testing of all IRR-manufactured reagents at bulk, post-fill and final production stages to confirm that reagents meet all defined acceptance criteria. Acceptance of noncomplying results will be at the discretion of the CDC COR, with technical input from CDC subject matter experts.

Test Results

Maintain Storage Cold Chain 100% acceptance of functional performance of QC samples after assembly.

Ability of shipping packaging to maintain reagents frozen must be validated prior to initial use, and on an annual basis thereafter.

18. Influenza Reagent Panels

Format Virtual Panels – Individual tubes bundled on demand, for immediate distribution. Panel components are also available individually (“a la carte”).

Made to order panels – Assembled into kits on demand, typically used for blinded panels.

Panel components may also be available individually (“a la carte”).

Physical panels – Assembled in advanced and stored in kit format. Panel components not available individually.

Container Virtual panel – vials shipped without additional outer packaging

Made to order panels – vials packed in a box or comparable secondary container, with an outer label applied just before shipment

Physical panels – vials packed in a mason box or comparable secondary container in advance, with an outer label applied before storage

Contractor may propose alternative packaging options for approval by CDC as needed.

Storage Storage conditions in accordance with specifications appropriate for that reagent type

Shipment Shipment conditions in accordance with specifications appropriate for that reagent type

Documentation Product Specification for each panel. Certificates of Analysis for each component lot produced.

Labeling All vial, box, and package insert proofs inspected and approved by CDC before printing.

CDC may remove individual panel components from a virtual panel or replace components with a more relevant strain in accordance with the quantities listed in Appendix A.

The Influenza Division will independently coordinate manufacture of reagents for the Influenza Molecular

Diagnostic Performance Evaluation Panel and Antiviral Resistance Panel. Reagents will be transferred to IRR for labeling, final kit assembly, storage, and distribution only.

Non-Influenza Respiratory Panel, Organisms Virtual Panel. Composed of 20-25 live viruses and 10-15 live bacteria reference strains.

Influenza Virus Isolation and Identification Panel Made to order panel. Contains 5 blinded influenza live virus strains and one negative control representing both

Influenza A and B types across a range HA titers.

Influenza Virus Isolation and Identification Panel, Supplemental Challenge Viruses

Made to order panel. Contains 2 blinded samples representing live influenza virus strains with challenging growth conditions.

Influenza Antigenicity Panel Virtual panel composed of 5 live viruses

Influenza Next Generation Performance Evaluation

Panel

Virtual panel composed of 10 sequencing grade live viruses

Influenza Next Generation Sequencing Limits of

Detection Panel

Physical panel composed of dilutions from 3 infectious virus stocks. Each stock should be diluted logarithmically for 8 serial dilutions (10-1 to 10-8).

Influenza Sequencing Panel Physical panel composed of four genomic RNA’s extracted from influenza viruses

Influenza Molecular Diagnostic Performance

Evaluation Panel

Physical panel manufactured by CDC. Contains 8-10 noninfectious (BPL inactivated) influenza to evaluate the performance of influenza real-time RT-PCR kits.

Antiviral Resistance Panel Physical panel manufactured by CDC. Contains 8 live viruses representing both Influenza A and B types. Viruses include both wild-type and antiviral-resistant or reduced inhibition variant viruses.

Quality Control Tests and Acceptance Criteria

CDC may perform independent verification QC testing of all IRR-manufactured reagents at bulk, post-fill and final production stages to confirm that reagents meet all defined acceptance criteria. Acceptance of noncomplying results will be at the discretion of the CDC COR, with technical input from CDC subject matter experts.

Test Results

All tests performed in accordance with specifications appropriate for that reagent type.

All results in accordance with specifications appropriate for that reagent type

19. Bacterial Pathogen RT-PCR Detection Kits

Format Kits containing lyophilized oligonucleotide primers and probes for the detection and identification of bacteria using RT-PCR technology:

(1) 1 forward primer, 1 reverse primer, and 1 probe set per kit

Kit includes one package insert

Container Fisher Nunc Transport Boxes for Cryoline, Fisher Scientific Cat # 12-565-184 inside clear resealable bag, with package insert folded so that labeling faces outward.

Contractor may propose alternative packaging options for approval by CDC as needed.

Storage 2 - 8°C

Shipment 2 - 8°C

Documentation Certificates of Analysis for each lot produced

Labeling All vial, box, and package insert proofs inspected and approved by CDC before printing.

CDC will provide primers/probes for comparing the functional performance of the first lot manufactured by IRR in order to establish initial assay performance.

Primers: 19-28 nucleotides

Probes: 22-45 nucleotides 5' mod

Internal mod 3' mod

Purification

(primers/probes)

Quantity per Vial

(forward, reverse, probe)

Streptococcus spp.

FAM BHQ-1 BHQ-1 RP HPLC/dual HPLC 7.5 nmol, 15 nmol, 5 nmol

Neisseria spp.

FAM BHQ-1

BHQ 1 or

Spacer 6 RP HPLC/dual HPLC 7.5 nmol, 7.5 nmol, 7.5 nmol

Haemophilus spp.

FAM BHQ-1 Spacer 6 RP HPLC/dual HPLC 2.5 nmol, 22 nmol, 7.5 nmol

Bordetella spp. FAM or

HEX or

Quas670

Spacer 6 or BHQ-1 or BHQ-3

RP HPLC or AX

HPLC/dual HPLC 5 nmol, 5 nmol, 4 nmol

Other bacterial pathogens or RNase P FAM BHQ-1 Spacer 6 RP HPLC/dual HPLC 15 nmol, 15 nmol, 7.5 nmol

* Preliminary configurations only, final production values may be adjusted after final assay optimization.

Quality Control Tests and Acceptance Criteria

CDC may perform independent QC testing of functional performance of all IRR-manufactured reagents at bulk, post-fill and final production stages to confirm that reagents meet all defined acceptance criteria. Acceptance of noncomplying results will be at the discretion of the CDC COR, with technical input from CDC subject matter experts.

Test Results

Sequence identity Verification of correct oligonucleotide sequence, including presence of all applicable sequence degeneracies

Sequence modifications Presence of correct fluorophore, quencher, and other modifications as applicable

Oligonucleotide purity ≥ 90% as determined by analytical HPLC or equivalent

Molecular weight ≤ 2.0% of theoretical molecular weight of the product including all applicable sequence degeneracies

Functional performance Specific detection of target pathogen within calibrated range for a given positive control (typically Cycle threshold between 22-27)

20. Atypical Respiratory Pathogens RT-PCR Detection Kits

Format Kits containing oligonucleotide primers and dual-labeled hydrolysis probe for the detection and identification of bacteria using RT-PCR technology:

(1) 4 primer/probe sets

Each set includes a forward primer, reverse primer and probe

Kit includes one package insert

Container Fisher Nunc Transport Boxes for Cryoline, Fisher Scientific Cat # 12-565-184 inside clear resealable bag, with package insert folded so that labeling faces outward.

Contractor may propose alternative packaging options for approval by CDC as needed.

Storage -20°C or colder

Shipment Frozen on dry ice

Documentation Certificates of Analysis for each lot produced

Labeling All vial, box, and package insert proofs inspected and approved by CDC before printing.

CDC will provide primers/probes for comparing the functional performance of the first lot manufactured by IRR in order to establish initial assay performance.

Primers: 18-23 nucleotides

Probes: 20-25 nucleotides 5' mod 3' mod

Purification

(primers/probes)

Quantity per Vial

(forward, reverse, probe)

Mycoplasma pneumonia HEX BHQ-1 Salt-Free/dual HPLC 2.5 nmol, 2.5 nmol, 1 nmol

Chlamydia pneumonia Quas670 BHQ-3 Salt-Free/dual HPLC 2.5 nmol, 2.5 nmol, 0.5 nmol

Legionella spp. FAM MGBNFQ Salt-Free/dual HPLC 1.25 nmol, 1.25 nmol, 0.25 nmol

RNase P CALRed610 BHQ-2 Salt-Free/dual HPLC 2.5 nmol, 2.5 nmol, 0.5 nmol

* Preliminary configurations only, final production values may be adjusted after final assay optimization.

Quality Control Tests and Acceptance Criteria

CDC may perform independent QC testing of functional performance of all IRR-manufactured reagents at bulk, post-fill and final production stages to confirm that reagents meet all defined acceptance criteria. Acceptance of noncomplying results will be at the discretion of the CDC COR, with technical input from CDC subject matter experts.

Test Results

Sequence identity Verification of correct oligonucleotide sequence, including presence of all applicable sequence degeneracies

Sequence modifications Presence of correct fluorophore, quencher, and other modifications as applicable

Oligonucleotide purity ≥ 90% as determined by analytical HPLC or equivalent

Molecular weight ≤ 2.0% of theoretical molecular weight of the product including all applicable sequence degeneracies

Functional performance Specific detection of target pathogen within calibrated range for a given positive

21. Streptococcus RT-PCR Typing Kit

Format Kit containing lyophilized oligonucleotide primers and probes for the typing of streptococcus bacteria using RT-PCR technology:

(1) 1 forward primer, 1 reverse primer, and 1 probe set per kit

Kit includes one package insert

Container Fisher Nunc Transport Boxes for Cryoline, Fisher Scientific Cat # 12-565-184 inside clear resealable bag, with package insert folded so that labeling faces outward.

Contractor may propose alternative packaging options for approval by CDC as needed.

Storage 2 to 8°C

Shipment 2 to 8°C

Documentation Certificates of Analysis for each lot…