CAJ | 학술논문

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 oxidase 1,CO1)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500 bp和700 bp的条带,2种PCR产物分别与美洲钩虫CO1(GenBank登录号为AF303136.1)和十二指肠钩虫CO1(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.
목적 대중국류행적미주구충화십이지장구충진행PCR감별.방법 통과대중국오성구충환자사용쌍간내산새밀정구충획득구충성충,추제단조미주구충화십이지장구충총DNA(각25조),용미주구충화십이지장구충선립체DNA세포색소양화매아기1(mitochondrial cytochrome oxidase 1,CO1)기인특이성인물(NaF-NaR,AdF-AdR)진행PCR확증.대PCR산물진행전영、측서.사용상동인물대일본혈흡충、편충、견구충DNA진행PCR확증.결과 25빈미주구충화25빈십이지장구충균능각확증출약500 bp화700 bp적조대,2충PCR산물분별여미주구충CO1(GenBank등록호위AF303136.1)화십이지장구충CO1(GenBank등록호위AJ417718.1)기인편단서렬일치성위99％화98％.단2대인물용우기타충충DNA칙무조대.결론 인물NaF-NaR、AdF-AdR능구용우구분재중국류행적미주구충화십이지장구충.
Objective To identify Necator americanus and Ancylostoma 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 Necator DNA by primer NaF-NaR,while 700 bp fragment was amplified by primer AdF-AdR from all 25 Ancylostoma 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.