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HCV Genotyping을 위한 염기서열분석법과 Restriction Fragment MassPolymorphism 법의 비교

The Comparison of Restriction Fragment Mass Polymorphism with Sequencing Method for the Hepatitis C Virus Genotyping

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참고문헌

주제분류병리학

초록

배경 : HCV유전형 검사는 간경화로의 진행이나 치료 반응도를 예측할 수 있는 지표이어서 매우 중요하다. 유전형 검사법으로 최근 개발되어 이용되는 restriction fragment mass polymorphism(RFMP)법은 혼합 감염시 유전형을 정확한 판정할 수 있다는 장점이 있다. 본 연구에서는 염기서열분석법과 비교하여 RFMP의 임상적 유용성을 평가하고자 하였다. 방법 : 서울아산병원에서 만성간질환으로 진단받은 43명의 환자를 대상으로 하였다. HCV유전자형은 RFMP와 염기서열분석법으로 결정하였다. 두 방법의 결과를 유전자형(genotype) 수준과 아형(subtype) 수준에서 일치율을 분석하였다. 결과 : 유전자형 수준에서 43예 중 43예가 모두(100%) 일치하였고 아형수준에서는 2예(4.7%)에서 불일치를 보였다. 불일치한 결과 2예는 유전자형 분석에서 1b형의 단일 감염으로 나타났으나 RFMP에서는 1b와 1c, 1b와 1a형으로 1b형을 포함한 혼합 감염으로 나타났다. 결론 : RFMP는 직접염기서열분석법과 비교하여 HCV유전자형 결정에 있어 우수한 일치율을 보여 유용한 방법이라고 사료되었다. 불일치한 결과는 아형 수준에서만 2예가 관찰되어 임상적으로 문제가 되지 않았다. 저자들의 연구결과 RFMP를 이용한 HCV 유전자형 결정은 매우 정확하며, 혼합 감염의 경우 직접염기서열 분석법보다 우수하다고 판단되었다.

Abstract

Background : Hepatitis C virus (HCV) genotyping is of increasing the importance in the progression to liver cirrhosis and the outcome of antiviral therapy. Restriction fragment mass polymorphism (RFMP) method which is developed recently for HCV genotyping is known to report accurate result in mixed infection. We evaluated the performance of RFMP in HCV genotyping by comparing the result of direct sequencing. Methods : Forty-three chronically HCV infected patients in Asan Medical Center were enrolled. HCV genotyping was performed by both RFMP and direct sequencing. The results were compared from the genotype level to the subtype level. Results : In the genotype level, all the results (100%) were concordant. At the subtype level, however, 2 results (2/43, 4.7%) were disconcordant. Two cases were reported as single infection of type 1b by direct sequencing while RFMP reported as mixed infection of 1b with 1a and 1b with 1c, respectively. Conclusions : Both sequencing and RFMP assays were highly concordant. We suggest that RFMP is a novel method in HCV genotyping superior to sequencing or hybridization method especially in mixed infection.

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