Establishment of a real-time fluorescent quantitative RT-PCR rapid detection method for human astrovirus / 中华微生物学和免疫学杂志

ABSTRACT

Objective:To establish a rapid detection method for human astrovirus based on TaqMan-probe real-time fluorescent quantitative reverse transcription-polymerase chain reaction (RT-PCR).Methods:According to the conservative sequence of human astrovirus ORF1 b gene, we designed the amplification primers and specific fluorescent probe to establish the human astrovirus TaqMan real-time fluorescent quantitative RT-PCR rapid detection method. The specificity, sensitivity and stability of the method were evaluated. We also used this method to detect human astrovirus in clinical samples. Results:The established human astrovirus TaqMan real-time fluorescent quantitative RT-PCR detection method has good specificity and repeatability for human astrovirus, and the sensitivity can reach 10 2 copies/μl. After testing the clinical samples, the detection rate of human astrovirus by our method was 100%. Conclusions:The human astrovirus TaqMan real-time fluorescent quantitative RT-PCR detection method established in this study is simple, rapid, sensitive, specific and stable. It can be used for clinical human astrovirus detection and epidemiological investigation.