Establishment of the 1st World Health Organization international standards for human papillomavirus type 16 DNA and type 18 DNA

• Published in: International journal of cancer 2010-06, Vol.126 (12), p.2969-2983
• Main Authors: Wilkinson, Dianna E., Baylis, Sally A., Padley, David, Heath, Alan B., Ferguson, Morag, Pagliusi, Sonia R., Quint, Wim G., Wheeler, Cosette M.
• Format: Article
• Language: English
• Subjects: assay standardization; Clinical Laboratory Techniques - standards; DNA, Viral - analysis; HPV DNA; Human papillomavirus; Human papillomavirus 16; Human papillomavirus 16 - genetics; Human papillomavirus 18 - genetics; Humans; International Cooperation; Nucleic Acid Amplification Techniques - standards; Polymerase Chain Reaction; Reference Standards; WHO international standards; World Health Organization
• ISSN: 0020-7136; 1097-0215
• DOI: 10.1002/ijc.25039

A World Health Organization collaborative study was conducted to evaluate candidate international standards for human papillomavirus (HPV) Type 16 DNA (NIBSC code 06/202) and HPV Type 18 DNA (NIBSC code 06/206) for use in the amplification and detection steps of nucleic acid‐based assays. The freeze‐dried candidate international standards were prepared from bulk preparations of cloned plasmid containing full‐length HPV‐16 or HPV‐18 genomic DNA. Nineteen laboratories from 13 countries participated in the study using a variety of commercial and in‐house quantitative and qualitative assays. The data presented here indicate that, upon freeze‐drying, there is no significant loss in potency for the candidate HPV‐18 DNA and a slight loss in potency for the candidate HPV‐16 DNA; although this is likely not scientifically relevant when assay precision is considered. In general, the individual laboratory mean estimates for each study sample were grouped ±∼2 log10 around the theoretical HPV DNA concentration of the reconstituted ampoule (1 × 107 HPV genome equivalents/mL). The agreement between laboratories is improved when potencies are made relative to the candidate international standards, demonstrating their utility in harmonizing amplification and detection steps of HPV‐16 and −18 DNA assays. Degradation studies indicate that the candidate international standards are extremely stable and suitable for long‐term use. Based on these findings, the candidate standards were established as the 1st WHO international standards for HPV‐16 DNA and HPV‐18 DNA, each with a potency of 5 × 106 international units (IU) per ampoule or 1 × 107 IU mL−1 when reconstituted as directed.