Sensitivity of PCR targeting the IS2404 insertion sequence of Mycobacterium ulcerans in an assay using punch biopsy specimens for diagnosis of buruli ulcer
Sensitivity of PCR targeting the IS2404 insertion sequence of Mycobacterium ulcerans in an assay using punch biopsy specimens for diagnosis of buruli ulcer
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DOI：
10.1128/jcm.43.8.3650-3656.2005
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发表时间：
2005-08-01
影响因子：
9.4
通讯作者：
Wansbrough-Jones, M
中科院分区：
文献类型：
--
作者：
Phillips, R;Horsfield, C;Wansbrough-Jones, M
对溃疡分枝杆菌病病灶穿刺活检标本进行了直接涂片、培养、PCR和组织病理学诊断的敏感性和特异性比较。溃疡病在现场设置。对插入元件IS 2404的PCR进行了修改，以包括尿嘧啶-N-糖基化酶和脱氧尿苷三磷酸，而不是脱氧胸苷三磷酸，以减少交叉污染的风险。临床诊断布鲁里溃疡的“金标准”是明确的组织学诊断，M培养阳性。溃疡，或抗酸杆菌（AFB）涂片阳性，以及可能的组织学诊断。对70例临床确诊的M.对溃疡病的敏感性达98%，且快速。显微镜、培养和组织学检查的敏感性分别为42%、49%和82%。使用4 mm穿刺活检标本优于6 mm穿刺活检标本，因为伤口不太可能出血和需要缝合。鉴于足够的技术专长和控制的使用，PCR在加纳的教学医院环境中是可行的;在常规实践中，我们建议使用活检标本的Ziehl-Neelsen染色来检测AFB，然后仅在AFB阴性病例中使用PCR，以最大限度地降低成本。组织学和培养仍然是重要的质量控制测试，特别是在治疗效果的研究。
Punch biopsy specimens from Mycobacterium ulcerans disease lesions were used to compare the sensitivities and specificities of direct smear, culture, PCR, and histopathology in making a diagnosis of M. ulcerans disease in a field setting. PCR for the insertion element IS2404 was modified to include uracil-N-glycosylase and deoxyuridine triphosphate instead of deoxythymidine triphosphate to reduce the risk of cross contamination. The "gold standard" for confirmation of clinically diagnosed Buruli ulcer was a definite histological diagnosis, a positive culture for M. ulcerans, or a smear positive for acid-fast bacilli (AFB), together with a possible histological diagnosis. For 70 clinically diagnosed cases of M. ulcerans disease, the modified PCR was 98% sensitive and gave a rapid result. The sensitivities of microscopy, culture, and histology were 42%, 49%, and 82%, respectively. The use of a 4-mm punch biopsy specimen was preferred to a 6-mm punch biopsy specimen since the wound was less likely to bleed and to need stitching. Given adequate technical expertise and the use of controls, the PCR was viable in a teaching hospital setting in Ghana; and in routine practice, we would recommend the use of Ziehl-Neelsen staining of biopsy specimens to detect AFB, followed by PCR, in AFB-negative cases only, in order to minimize costs. Histology and culture remain important as quality control tests, particularly in studies of treatment efficacy.