【摘要】 目的:明确单管巢式实时PCR检测麻风杆菌的特异性、灵敏性和重复性。方法:以麻风杆菌hsp18基因为靶基因设计引物和探针,建立单管巢式实时PCR方法检测麻风杆菌并与抗酸染色及普通PCR进行特异性和灵敏性的比较。结果:单管巢式实时PCR检测下限为2.1fg质粒DNA,普通PCR为21fg质粒DNA,敏感性高10倍;与8种非麻风杆菌无交叉反应;批内相对标准偏差(RSD)为0.24%~1.08%,批间RSD为0.09%~2.8%。对比检测54例麻风疑似标本中,抗酸染色、普通PCR和单管巢式实时PCR的敏感性分别为81.3%(26/32)、87.5%(28/32)和96.9%(31/32)。结论:单管巢式实时PCR特异性强、敏感性高且重复性好。更多还原

【Abstract】 Objective;To detect the sensitivity of single-tube nested realtime PCR in detecting Mycobacterium leprae.Methods;The primers and probe were designed based on the hsp18 gene of Mycobacterium leprae.The single-tube nested realtime PCR was established and the sensitivity and specificity were compared with acid-fast staining and ordinary PCR.Results:The sensitivity of the single-tube nested realtime PCR for detecting Mycobacterium leprae was 2.1fg of plasmid DNA,which was 10 times more sensitive than that in the normal PCR.There was non-cross reactivity with other 8 Non-Mycobacterium leprae.Intra relative standard deviation(RSD) was 0.24%-1.08%and inter RSD was 0.09%-2.8%.The sensitivity of acid-fast staining,normal and single-tube nested PCR in detecting 54 suspected samples of leprosy was 81.3%(26/32),87.5%(28/32) and 96.9%(31/32),respectively.Conclusion;The single-tube nested realtime PCR method in detecting Mycobacterium leprae is more sensitive and specific.更多还原