PCR identification of two hookworm species in China

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. Key words: Hookworm; Molecular marker; Identification