임상 가검물에서 Nested-PCR 과 PCR - dot blot 법을 이용한 결핵균 검출의 비교

Abstract

The polymerase chain reaction (PCR) technique has widely been used in the direct diagnosis of Mycobacterium tuberculosis(M. tuberculosis) from variety of clinical samples. Recently, with development of molecular biological techniques, several PCR methods, including nested - PCR, PCR- RFLP(Restriction Fragment Length Polymorphism) and PCR-dot blot hybridization, have been applied for the clinical diagnosis of M. tubercul osis. In this study, we evaluated the sensitivity of nested - PCR in comparison with the PCR-dot blot for the determination of M. tuberculosis. Two pairs of primers for nested - PCR were designed from IS6110 locus of M. tuberculosis. Biotin labeled probe designed in the inner area of PCR product was used for PCR-dot hybridization. In a total of 59 sputum samples, thirty five samples (59.3 %) were positive in nested - PCR. On the other hand, fifteen samples (25.4 %) were positive by PCR - dot blot analysis. From the results, nested - PCR is a more simple , cost effective and sensitive method for the detec tion of M. tuberculosis than PCR - dot blot analysis.