Molecular detection and identification of Trichobilharzia: development of a LAMP, qPCR, and multiplex PCR toolkit

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216208%3A11310%2F25%3A10499183" target="_blank" >RIV/00216208:11310/25:10499183 - isvavai.cz</a>

Result on the web

<a href="https://verso.is.cuni.cz/pub/verso.fpl?fname=obd_publikace_handle&handle=FzWOCmaERd" target="_blank" >https://verso.is.cuni.cz/pub/verso.fpl?fname=obd_publikace_handle&handle=FzWOCmaERd</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1186/s13071-025-06822-y" target="_blank" >10.1186/s13071-025-06822-y</a>

Result language

angličtina

Original language name

Molecular detection and identification of Trichobilharzia: development of a LAMP, qPCR, and multiplex PCR toolkit

Original language description

Background: Cercarial dermatitis (CD), or swimmer&apos;s itch, is a water-borne allergic skin reaction caused by the penetration of the larval stages of bird schistosomes (cercariae) into the skin. Members of the genus Trichobilharzia are the primary causative agents of CD worldwide. Due to the increasing number of cases, CD is regarded as a (re)emerging disease. Outbreaks in recreational waters can significantly impact public health and local economies. Environmental monitoring of Trichobilharzia is crucial for outbreak prediction and public health management. However, conventional methods, such as cercarial shedding and snail dissections, are labour-intensive and lack sensitivity. To overcome these limitations, we present a molecular toolkit that combines loop-mediated isothermal amplification (LAMP), quantitative polymerase chain reaction (qPCR), and multiplex PCR for rapid, sensitive, and accurate detection and identification of Trichobilharzia spp. from various biological samples.Methods: Tricho-LAMP and Tricho-qPCR were designed and optimised for Trichobilharzia DNA detection. A multiplex PCR assay was also developed and optimised to identify the three main species causing CD in Europe (Trichobilharzia franki, T. szidati, and T. regenti).Results: Tricho-LAMP specifically detected T. regenti and T. franki at 10(-3) ng, and T. szidati at 10(-2) ng per reaction with genomic DNA. Using gBlocks synthetic DNA, Tricho-LAMP achieved 100% amplification at 10,000 copies and 85% amplification at 1000 copies, with decreasing success at lower concentrations. Tricho-qPCR showed the highest sensitivity, detecting all species down to 10(-4) ng per reaction and showing a limit of detection at 10 copies of synthetic DNA in the reaction. Multiplex PCR allowed reliable species differentiation via gel electrophoresis of the PCR products, but the assay had the lowest sensitivity.Conclusions: We provide a molecular toolkit consisting of LAMP, qPCR, and multiplex PCR. By exhibiting high sensitivity, Tricho-LAMP and Tricho-qPCR assays are potentially suitable for environmental DNA (eDNA)-based environmental monitoring of bird schistosomes, by both researchers and public health authorities. Multiplex PCR can be used for species determination without the need for further sequencing.

Czech name

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Czech description

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Type

J_imp - Article in a specialist periodical, which is included in the Web of Science database

CEP classification

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OECD FORD branch

10600 - Biological sciences

Project

—

Continuities

S - Specificky vyzkum na vysokych skolach<br>I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Publication year

2025

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Name of the periodical

Parasites &amp; Vectors

ISSN

1756-3305

e-ISSN

1756-3305

Volume of the periodical

18

Issue of the periodical within the volume

1

Country of publishing house

GB - UNITED KINGDOM

Number of pages

11

Pages from-to

195

UT code for WoS article

001499223400004

EID of the result in the Scopus database

2-s2.0-105006909275