Yamasaki Hiroshi, Allan James C, Sato Marcello Otake, Nakao Minoru, Sako Yasuhito, Nakaya Kazuhiro, Qiu Dongchuan, Mamuti Wulamu, Craig Philip S, Ito Akira
Department of Parasitology. Animal Laboratory for Medical Research, Asahikawa Medical College, Asahikawa, Japan.
J Clin Microbiol. 2004 Feb;42(2):548-53. doi: 10.1128/JCM.42.2.548-553.2004.
Multiplex PCR was established for differential diagnosis of taeniasis and cysticercosis, including their causative agents. For identification of the parasites, multiplex PCR with cytochrome c oxidase subunit 1 gene yielded evident differential products unique for Taenia saginata and Taenia asiatica and for American/African and Asian genotypes of Taenia solium with molecular sizes of 827, 269, 720, and 984 bp, respectively. In the PCR-based detection of tapeworm carriers using fecal samples, the diagnostic markers were detected from 7 of 14 and 4 of 9 T. solium carriers from Guatemala and Indonesia, respectively. Test sensitivity may have been reduced by the length of time (up to 12 years) that samples were stored and/or small sample volumes (ca. 30 to 50 mg). However, the diagnostic markers were detected by nested PCR in five worm carriers from Guatemalan cases that were found to be negative by multiplex PCR. It was noteworthy that a 720 bp-diagnostic marker was detected from a T. solium carrier who was egg-free, implying that it is possible to detect worm carriers and treat before mature gravid proglottids are discharged. In contrast to T. solium carriers, 827-bp markers were detected by multiplex PCR in all T. saginata carriers. The application of the multiplex PCR would be useful not only for surveillance of taeniasis and cysticercosis control but also for the molecular epidemiological survey of these cestode infections.
建立了多重PCR用于绦虫病和囊尾蚴病的鉴别诊断，包括其病原体。为了鉴定寄生虫，使用细胞色素c氧化酶亚基1基因的多重PCR产生了明显的差异产物，分别为牛肉绦虫、亚洲绦虫以及猪带绦虫美洲/非洲和亚洲基因型所特有，分子大小分别为827、269、720和984 bp。在基于PCR的使用粪便样本检测绦虫携带者的过程中，分别从危地马拉和印度尼西亚的14名猪带绦虫携带者中的7名以及9名中的4名检测到了诊断标记。检测灵敏度可能因样本储存时间（长达12年）和/或样本量小（约30至50毫克）而降低。然而，在多重PCR检测为阴性的危地马拉病例中的5名绦虫携带者中，通过巢式PCR检测到了诊断标记。值得注意的是，在一名无虫卵的猪带绦虫携带者中检测到了720 bp的诊断标记，这意味着有可能在成熟孕节排出之前检测到绦虫携带者并进行治疗。与猪带绦虫携带者不同，在所有牛肉绦虫携带者中通过多重PCR检测到了827 bp的标记。多重PCR的应用不仅对绦虫病和囊尾蚴病控制的监测有用，而且对这些绦虫感染的分子流行病学调查也有用。
