뇌척수액에서 PCR-RFLP법을 이용한 Herpes Simpiex Virus Ⅰ, Ⅱ의 동정에 관한 연구

Abstract

Three different DNA extraction methods including phenol/chloroform, GuSCN/silica and boiling methods were tried to increased sensitivities of detection of HSV by PCR in clinical CSFs. And types of HSV were identified by Polymerase Chain Reaction- Restriction Gragment Length Polymorphism(PCR-RFLP). Also these samples were analysed by Enzyme Immuno Assay(EIA) to compare with PCR results. 476 bp of amplified DNA products was restricted by AvaII and resulted in 206, 183, 87 fragment for HSV I, 389 and 87 fragment for HSV II, respectively. In a total of 58 CSF samples, all samples were IgM and IgG negative by EIA. However 8 samples were positive by Pcr. And by PCR-RFLP, HSV type I was 6 cases, HSV type II was 2 cases. In 6 HSV I, phenol and silica extraction methid were same result as 6 case. But 5 were positive in boiling method. In 2 HSV II, phenol and silica extraction method were also same results as 2 cases. But one was positive in boiling method. AS above our results, phenol and silica estraction method were usefullness for detection of HSV in CSF by PCR. Also PCR was more sensitive as 13.8% then EIA. In conclusion, HSV PCR-RFLP method provides relaticcely rapid and practical tool in CSF samples for the early diagnosis of HSV in fection.