Studies on Detecting Brugia malayi Larva in Mosquitoes by Polymerase Chain Reaction / 中国寄生虫学与寄生虫病杂志

ABSTRACT

Objective To establish a specific, sensitive and simple assay for the detection of Brugia malayi larva in Anopheles sinensis .Methods Using a new DNA purification technique (Microcon 100) and two pairs of oligonucleotide primers (p1, p2 and p3,p4) suitable for detecting B malayi in seven areas in our country, the mosquito vectors infected by B malayi were detected by polymerase chain reaction(PCR).Results This PCR method could amplify separately a 322 basepair(bp) and a 155 bp DNA fragment and detect as few as 1/64 of one L 1 in 1 mosquito,the detectable limit was nearly 4 pg DNA of filarial larvae, and it could also detect 1 infected mosquito with one L 3 of B malayi in pools of up to 200 mosquitoes. In contrast,no such specific 322 bp or 155 bp DNA band was detected in Dilofilaria immitis and normal mosquito.Conclusion This PCR techique established for supervision of mosquito vector in B malayi endemic areas is specific,sensitive,and simple.