Detection of hantavirus by RT - nested PCR in patients with hemorrhagic fever with renal syn- drome

Hantavirus (HV) has a tripartite, single - stranded, negative - sense RNA genome. Two pairs of oligonucleotides flanking S segment were chosen as primers for reverse transcription Polymerase chain reaction(RT - nested PCR). The method of RT - nested PCR was used for detection of virus RNA in HV infected Vero E6 cells and clinical serum specimens from 35 hemorrhagic fever with renal syndrome(HFRS) patients, with HV gene S segment plasmid cDNA as postive control,and virus RNA was extracted by Acid Guanidinium Thiocyanate - Phenol - Chloroform Single- Step Method. The results showed that all the patients' serum samples and HV infected Vero E6 cells were positive, on the other hand, all the twenty normal serum samples and Vero E6 cells were negative. It suggested that RT - nested PCR was sensitive, specific, rapid and simple, and can be applied for study on the pathogenesis of HFRS in molecular level and early clinical diagnosis