Sensitive and rapid detection of Paragonimus westermani infection in humans and animals by loop-mediated isothermal amplification (LAMP)

被引：38

作者：

Chen, M. X. ^{[2
]}

Ai, L. ^{[2
,3
]}

Zhang, R. L. ^{[4
]}

Xia, J. J. ^{[5
]}

Wang, K. ^{[5
]}

Chen, S. H. ^{[2
]}

Zhang, Y. N. ^{[2
]}

Xu, M. J. ^{[1
]}

Li, X. ^{[3
]}

Zhu, X. Q. ^{[1
]}

Chen, J. X. ^{[2
]}

机构：

[1] Chinese Acad Agr Sci, Lanzhou Vet Res Inst, State Key Lab Vet Etiol Biol, Key Lab Vet Parasitol Gansu Prov, Lanzhou 730046, Gansu, Peoples R China

[2] Chinese Ctr Dis Control & Prevent, Natl Inst Parasit Dis, Shanghai 200025, Peoples R China

[3] S China Agr Univ, Coll Vet Med, Guangzhou 510642, Guangdong, Peoples R China

[4] Shenzhen Ctr Dis Control & Prevent, Shenzhen 518020, Guangdong, Peoples R China

[5] Shenzhen Med Continuing Educ Ctr, Shenzhen 518020, Guangdong, Peoples R China

来源：

PARASITOLOGY RESEARCH | 2011年 / 108卷 / 05期

关键词：

MOLECULAR-CLONING; PCR; IDENTIFICATION; METACERCARIAE; DIAGNOSIS; CRABS;

D O I：

10.1007/s00436-010-2162-x

中图分类号：

R38 [医学寄生虫学]; Q [生物科学];

学科分类号：

07 ; 0710 ; 09 ; 100103 ;

摘要：

In the present study, a loop-mediated isothermal amplification (LAMP) assay was developed and validated for the detection of Paragonimus westermani adults, metacercariae, and eggs in human and animal samples. The LAMP amplification can be finished in 45 min under isothermal condition at 60A degrees C by employing a set of four species-specific primer mixtures and the results can be checked by naked-eye visualization. No amplification products were detected with deoxyribunucleic acid (DNA) of related trematode species including Fasciola hepatica, Fasciola gigantica, Clonorchis sinensis, Opisthorchis viverrini, Schistosoma mansoni, and Schistosoma japonicum. The method was further validated by examining P. westermani DNA in intermediate hosts including freshwater crabs and crayfish, as well as in sputum and pleural fluid samples from patients of paragonimiasis. These results indicated that the LAMP assay was highly specific, sensitive, and rapid, and it was approximately 100 times more sensitive than conventional specific PCR. The LAMP assay established in this study provides a rapid and sensitive tool for the detection of P. westermani DNA in freshwater crabs, crayfish, sputum, and pleural fluid samples, which has important implications for effective control of human paragonimiasis.

引用

页码：1193 / 1198

页数：6

共 33 条

[11] A seroepidemiological survey for paragonimosis among boar-hunting dogs in central and southern Kyushu, Japan [J].