Fischer P, Rubaale T, Meredith S E, Büttner D W
Bernhard Nocht Institute for Tropical Medicine, Hamburg, Germany.
Parasitol Res. 1996;82(5):395-401. doi: 10.1007/s004360050135.
A polymerase chain reaction (PCR) of a 150-bp tandem repeat of Onchocerca volvulus (O-150) combined with Southern-blot hybridization to species-specific DNA probes was employed for DNA detection. O-150 was amplified from parasites originating from Uganda, Benin, Cameroon, Liberia, Ghana, Burkina Faso, Mali, and Zaire and was successfully hybridized to digoxigenin-labeled oligonucleotides. To investigate the sensitivity of the PCR, 2 skin biopsies were taken from each of 227 persons from Uganda with proven O. volvulus infections but with low microfilaria (mf) densities due to ivermectin treatment. One biopsy was tested by PCR and the other was digested using collagenase to assess the total number of mf. The PCR revealed 76.2% of the samples to be positive, and the collagenase method showed that 78.9% were positive, indicating similar sensitivity for the two methods. It is probable that for both techniques the biopsy must contain at least one live mf or fragments of a dead mf. In this study, no free or circulating O. volvulus DNA could be detected in skin biopsies by PCR.
采用盘尾丝虫150碱基对串联重复序列(O-150)的聚合酶链反应(PCR)并结合物种特异性DNA探针的Southern印迹杂交进行DNA检测。从源自乌干达、贝宁、喀麦隆、利比里亚、加纳、布基纳法索、马里和扎伊尔的寄生虫中扩增出O-150,并成功与地高辛标记的寡核苷酸杂交。为研究PCR的敏感性,从227名来自乌干达且经证实感染盘尾丝虫但因伊维菌素治疗而微丝蚴(mf)密度较低的人中,每人取2份皮肤活检样本。一份活检样本用PCR检测,另一份用胶原酶消化以评估mf总数。PCR显示76.2%的样本呈阳性,胶原酶法显示78.9%呈阳性,表明两种方法的敏感性相似。两种技术可能都要求活检样本中至少含有一条活的mf或死mf的片段。在本研究中,通过PCR未在皮肤活检样本中检测到游离或循环的盘尾丝虫DNA。