摘要
目的对中国流行的美洲钩虫和十二指肠钩虫进行PCR鉴别。方法通过对中国五省钩虫患者使用双羟萘酸噻嘧啶驱虫获得钩虫成虫，抽提单条美洲钩虫和十二指肠钩虫总DNA（各25条），用美洲钩虫和十二指肠钩虫线粒体DNA细胞色素氧化酶亚基1（mitochondrial cytochrome oxidase1，C01）基因特异性引物（NaF—NaR，AdF—AdR）进行PCR扩增。对PCR产物进行电泳、测序。使用相同引物对日本血吸虫、鞭虫、犬钩虫DNA进行PCR扩增。结果25份美洲钩虫和25份十二指肠钩虫均能各扩增出约500bp和700bp的条带，2种PCR产物分别与美洲钩虫C01（GenBank登录号为AF303136．1）和十二指肠钩虫C01（GenBank登录号为AJ417718．1）基因片段序列一致性为99％和98％。但2对引物用于其他虫种DNA则无条带。结论引物NaF—NaR、AdF—AdR能够用于区分在中国流行的美洲钩虫和十二指肠钩虫。
Objective To identify- Neeator americanas and Aneylostoma duodenale in China by PCR. Methods Adult hookworms were collected through treating patients in 5 provinces of China with Pyrantel Pamoate. Total DNA from 25 Necators and 25 A. duodenales was extracted separately. Specific primers （ NaF- NaR for Nacator and AdF-AdR for A. duodenale） according to cytochrome c oxidase subunit 1 gene were used for PCR amplification. The products were analyzed by electrophoresis and sequencing. DNA of Schistosoma japomicum, Trichuris trichiura, Ancylostoma caninum were also tested with the same primers for PCR. Results 500 bp fragment was amplified from all 25 Neeator DNA by primer NaF-NaR, while 700 bp fragment was amplified by primer AdF-AdR from all 25 Aneylostoma duodenale DNA. Sequence alignment analysis showed that the two PCR products had 98% consistency with N. americanus CO1 （GenBank Accession No. AF303136.1 ） and A. duodenale CO1 （GenBank Accession No. AJ417718.1 ）. No band was shown when the same primer was used on other helminthes. Conclusion Primer NaF-NaR, AdF-AdR could be used to identify N. americanus and A. duodenale in China.
出处
《国际医学寄生虫病杂志》
CAS
2012年第2期94-97,共4页
International JOurnal of Medical Parasitic Diseases
基金
中国疾病预防控制中心寄生虫病预防控制所紧缺人才专项基金（2010）
中国热带病创新药物及诊断试剂国际合作研究（2010DFA33970）
关键词
钩虫
分子标记物
鉴别
Hookworm
Molecular marker
Identification
