Preparation of the first Chinese national standard substance of reverse transcriptase for activity assays / 中国药学杂志

ABSTRACT

OBJECTIVE: To establish the first national standard substance of reverse transcriptase for activity assays. METHODS: The standard substance of reverse transcriptase was prepared and tested according to the requirements in Chinese Pharmacopoeia (Volume III, 2015 edition). The stability of the standard substance was evaluated by thermal acceleration stability test. The activity was calibrated collaboratively by four laboratories using SYBR GreenIreal-time quantitative PCR method. RESULTS: The prepared standard substance was qualified in appearance and sterility tests. The moisture content was 1.3% and the dispensing accuracy was 0.75%. Thermal acceleration stability test showed that the enzyme activity of the standard substance stored at -20, 4, 25 and 37℃ for 3 and 6 months was not decreased compared with that stored at -70℃. Twenty-two calibration tests by four laboratories showed that the geometric mean enzyme activity of the standard was 26 U·vial-1, with a 95% confidence interval of 22.95-30.45 U·vial-1. CONCLUSION: The candidate standard substance of reverse transcriptase is judged to be suitable to serve as a Chinese national standard for its activity assays, and the enzyme activity specification is established to be 26 U·vial-1.