Newly developed SYBR Green-based quantitative real-time PCRs revealed coinfection evidence of Angiostrongylus cantonensis and A. malaysiensis in Achatina fulica existing in Bangkok Metropolitan, Thailand

• DOI: 10.1016/j.fawpar.2021.e00119
• Publication Date: 2021-03-19T00:29:22Z
• Authors: Wallop Jakkul; Kittipong Chaisiri; Naowarat Saralamba; Yanin Limpanont; Sirilak Dusitsitt...; Vachirapong Charo...; Abigail Hui En Chan; Urusa Thaenkham
• Keywords: Angiostrongyliasis; Achatina

Angiostrongylus cantonensis is a well-known pathogen causing eosinophilic meningitis associated with angiostrongyliasis. Humans, as accidental hosts, are infected by consuming undercooked snails containing third-stage larvae. A. malaysiensis closely related to and has been described potential human pathogen. The two species distribution was recently reported overlap in the same endemic area, particularly Indochina Peninsula. Similar morphological characteristics of larva snail-intermediate host often lead misidentification species. Thus, we aimed develop sensitive specific method detect discriminate designing species-specific primers based on mitochondrial cytochrome b gene. We developed SYBR Green quantitative real-time PCR (qPCR) for detection assays, which could be conducted simultaneously. subsequently employed identify larvae isolated from Achatina fulica collected six public parks Bangkok Metropolitan, Thailand. also preliminary applied parasite tissue debris patients' cerebrospinal fluid (CSF). qPCRs quantitatively detected approximately 10−4 ng genomic DNA one larva, facilitating detection. Based pools individually each parks, qPCR results revealed that predominant infecting 5.26% snails. In comparison, coinfection between 5.97%, no single infection fulica. Our useful inexpensive technique discrimination, sufficient sensitivity host. ratio can estimated molecular survey within intermediate hosts clinical diagnosis angiostrongyliasis CSF specimens future studies.