Evaluation of Loop-Mediated Isothermal Amplification Suitable for Molecular Monitoring of Schistosome-Infected Snails in Field Laboratories

被引:77
作者
Hamburger, Joseph
Abbasi, Ibrahim
Kariuki, Curtis [1 ]
Wanjala, Atsabina [1 ]
Mzungu, Elton [1 ]
Mungai, Peter [1 ]
Muchiri, Eric [1 ]
King, Charles H. [2 ]
机构
[1] Minist Publ Hlth & Sanitat, Div Vector Borne & Neglected Trop Dis, Nairobi, Kenya
[2] Case Western Reserve Univ, Sch Med, Ctr Global Hlth & Dis, Cleveland, OH 44106 USA
基金
美国国家卫生研究院;
关键词
CHAIN-REACTION ASSAY; EARLY PREPATENCY; HAEMATOBIUM DNA; COASTAL KENYA; TRANSMISSION; PCR; SURVEILLANCE; JAPONICUM; SEQUENCE; MANSONI;
D O I
10.4269/ajtmh.2012.12-0208
中图分类号
R1 [预防医学、卫生学];
学科分类号
1004 ; 120402 ;
摘要
We previously described loop-mediated isothermal amplification (LAMP) for detection of Schistosoma haematobium and S. mansoni DNA in infected snails. In the present study, we adapted the LAMP assay for application in field laboratories in schistosomiasis-endemic areas. Isolation of DNA was simplified by blotting snail tissue (extracted in NaOH/sodium dodecyl sulfate) onto treated membranes, which enabled preservation at ambient temperatures. A ready-mix of LAMP reagents, suitable for shipment at ambient temperature and storage in minimal refrigeration, was used. Local survey teams without experience in molecular biology acquired operational expertise with this test within a few hours. Fifty-four field-caught snails were tested locally by LAMP and 59 were tested at similar conditions in Jerusalem. The LAMP results were consistent with those of a polymerase chain reaction; only four samples showed false-negative results. Results indicate that LAMP assays are suitable for detection of S. haematobium and S. mansoni in low-technology parasitology laboratories in which schistosomiasis elimination activities are undertaken.
引用
收藏
页码:344 / 351
页数:8
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