Loop-mediated isothermal amplification: rapid detection of Angiostrongylus cantonensis infection in Pomacea canaliculata.
BACKGROUND
Angiostrongylus cantonensis is a zoonotic parasite that causes eosinophilic meningitis in humans. The most common source of infection with A. cantonensis is the consumption of raw or undercooked mollusks (e.g., snails and slugs) harbouring infectious third-stage larvae (L3). However, the parasite is difficult to identify in snails. The purpose of this study was to develop a quick, simple molecular method to survey for A. cantonensis in intermediate host snails.


RESULTS
We used a loop-mediated isothermal amplification (LAMP) assay, which was performed using Bst DNA polymerase. Reactions amplified the A. cantonensis 18S rRNA gene and demonstrated high sensitivity; as little as 1 fg of DNA was detected in the samples. Furthermore, no cross-reactivity was found with other parasites such as Toxoplasma gondii, Plasmodium falciparum, Schistosoma japonicum, Clonorchis sinensis, Paragonimus westermani and Anisakis. Pomacea canaliculata snails were exposed to A. cantonensis first-stage larvae (L1) in the laboratory, and L3 were observed in the snails thirty-five days after infection. All nine samples were positive as determined by the LAMP assay for A. cantonensis, which was identified as positive by using PCR and microscopy, this demonstrates that LAMP is sensitive and effective for diagnosis.


CONCLUSIONS
LAMP is an appropriate diagnostic method for the routine identification of A. cantonensis within its intermediate host snail P. canaliculata because of its simplicity, sensitivity, and specificity. It holds great promise as a useful monitoring tool for A. cantonensis in endemic regions.
Institute of Parasitology, Zhejiang Academy of Medical Sciences, Hangzhou, People's Republic of China.
Rui
R
Chen
QunBo
Q
Tong
Yi
Y
Zhang
Di
D
Lou
QingMing
Q
Kong
Shan
S
Lv
MingMing
M
Zhuo
LiYong
L
Wen
ShaoHong
S
Lu
D017185:Angiostrongylus cantonensis / Q000145:classification / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D000818:Animals; D004197:Disease Reservoirs / Q000469:parasitology*; D004199:Disease Vectors; D006801:Humans; D007814:Larva / Q000235:genetics / Q000254:growth & development; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012680:Sensitivity and Specificity; D012908:Snails / Q000469:parasitology*; D017206:Strongylida Infections / Q000469:parasitology* / Q000635:transmission