Chansiri Kosum, Phantana Sirichai
Department of Biochemistry, Faculty of Medicine, Srinakharinwirot University, Bangkok, Thailand.
Southeast Asian J Trop Med Public Health. 2002 Sep;33(3):504-8.
A polymerase chain reaction (PCR) assay based on a highly repeated DNA sequence found in Wuchereria bancrofti (SspI repeat) has been modified for the survey of bancroftian filariasis in expatriate workers (Myamese, Karen and Mon) from Myanmar where human filariasis is endemic. The PCR was very sensitive with the ability to detect the presence of as little as 10 pg of parasite DNA. The primers used in this PCR also showed highly specific amplification of parasite DNA without the presence of non-specific and non-target PCR products such as Brugia malayi, Plasmodium falciparum and human DNA. The primers were used to investigate filariasis in four provinces in the central and western Thailand, Samut Songkram, Ratchaburi, Nakhon Pathom and Tak during 1997-2001. Among them, Tak and Ratchaburi are the only endemic areas of bancroftian filariasis. In this field study, 1,299 human blood samples (501 from Samut Songkram, 510 from Ratchaburi, 109 from Nakhon Pathom, and 179 from Tak) were collected and screened by PCR. The result showed that 1, 2, 3, and 33 patients from Samut Songkram, Ratchaburi, Nakhon Pathom, and Tak respectively were infected with W. bancrofti. These numbers were corresponded to the prevalence rate of infection of 0.2, 0.4, 2.8, and 18.5%, respectively. The PCR was able to detect the third-stage infectious larvae (L3) from Culex quinquefasciatus, mosquito vector of the W. bancrofti, that was experimentally fed to infected patient. The PCR screening of each of field mosquito pools from two endemic areas, Ratchaburi and Tak, showed that no L3 of W. bancrofti was detected.
基于在班氏吴策线虫中发现的高度重复DNA序列(SspI重复序列)的聚合酶链反应(PCR)检测方法已被改进,用于调查来自缅甸(人类丝虫病流行地区)的外籍工人(缅族、克伦族和孟族)中的班氏丝虫病。该PCR检测非常灵敏,能够检测到低至10 pg的寄生虫DNA。此PCR中使用的引物还显示出对寄生虫DNA的高度特异性扩增,不存在诸如马来布鲁线虫、恶性疟原虫和人类DNA等非特异性和非靶向PCR产物。在1997 - 2001年期间,这些引物被用于调查泰国中部和西部四个省份(巴蜀府、叻丕府、佛统府和夜丰颂府)的丝虫病情况。其中,夜丰颂府和叻丕府是班氏丝虫病仅有的流行地区。在这项实地研究中,收集了1299份人类血液样本(501份来自巴蜀府,510份来自叻丕府,109份来自佛统府,179份来自夜丰颂府),并通过PCR进行筛查。结果显示,巴蜀府、叻丕府、佛统府和夜丰颂府分别有1例、2例、3例和33例患者感染了班氏吴策线虫。这些数字分别对应0.2%、0.4%、2.8%和18.5%的感染患病率。该PCR能够检测到来自班氏吴策线虫的蚊媒——致倦库蚊经实验性喂食感染患者后体内的第三期感染性幼虫(L3)。对来自两个流行地区(叻丕府和夜丰颂府)的野外蚊虫池进行PCR筛查,结果显示未检测到班氏吴策线虫的L3。