Real-time fluorescence loop-mediated isothermal amplification assays for detection of zoonotic malaria Plasmodium parasites.

Lai, Meng Yee; Abdullah, Mohd Lutfi; Lau, Yee Ling

Lai, Meng Yee; Abdullah, Mohd Lutfi; Lau, Yee Ling.

Afiliación

Lai MY; Department of Parasitology, Faculty of Medicine, Universiti Malaya, 50603 Kuala Lumpur, Malaysia.
Abdullah ML; National Wildlife Forensic Laboratory, Ex-Situ Conservation Division Department of Wildlife and National Parks Peninsular Malaysia, Jalan Cheras, 56100 Kuala Lumpur, Malaysia.
Lau YL; Department of Parasitology, Faculty of Medicine, Universiti Malaya, 50603 Kuala Lumpur, Malaysia. Electronic address: lauyeeling@um.edu.my.

Acta Trop ; 255: 107249, 2024 Jul.

Article en En | MEDLINE | ID: mdl-38740319

ABSTRACT

ABSTRACT

BACKGROUND:

Natural human infections by Plasmodium cynomolgi and P. inui have been reported recently and gain the substantial attention from Southeast Asian countries. Zoonotic transmission of non-human malaria parasites to humans from macaque monkeys occurred through the bites of the infected mosquitoes. The objective of this study is to establish real-time fluorescence loop-mediated isothermal amplification (LAMP) assays for the detection of zoonotic malaria parasites by combining real-time fluorescent technology with the isothermal amplification technique.

METHODS:

By using 18S rRNA as the target gene, the primers for P. cynomolgi, P. coatneyi and P. inui were newly designed in the present study. Four novel real-time fluorescence LAMP assays were developed for the detection of P. cynomolgi, P. coatneyi, P. inui and P. knowlesi. The entire amplification process was completed in 60 min, with the assays performed at 65 °C. By using SYTO-9 as the nucleic acid intercalating dye, the reaction was monitored via real-time fluorescence signal.

RESULTS:

There was no observed cross-reactivity among the primers from different species. All 70 field-collected monkey samples were successfully amplified by real-time fluorescence LAMP assays. The detection limit for P. cynomolgi, P. coatneyi and P. knowlesi was 5 × 109 copies/µL. Meanwhile, the detection limit of P. inui was 5 × 1010 copies/µL.

CONCLUSION:

This is the first report of the detection of four zoonotic malaria parasites by real-time fluorescence LAMP approaches. It is an effective, rapid and simple-to-use technique. This presented platform exhibits considerable potential as an alternative detection for zoonotic malaria parasites.

Asunto(s)

Malaria; Técnicas de Diagnóstico Molecular; Técnicas de Amplificación de Ácido Nucleico; Plasmodium; ARN Ribosómico 18S; Sensibilidad y Especificidad; Zoonosis; Animales; Técnicas de Amplificación de Ácido Nucleico/métodos; Malaria/diagnóstico; Malaria/parasitología; Malaria/veterinaria; ARN Ribosómico 18S/genética; Técnicas de Diagnóstico Molecular/métodos; Plasmodium/genética; Plasmodium/aislamiento & purificación; Plasmodium/clasificación; Zoonosis/parasitología; Zoonosis/diagnóstico; Humanos; Cartilla de ADN/genética; Fluorescencia; Macaca/parasitología; Enfermedades de los Monos/parasitología; Enfermedades de los Monos/diagnóstico

Palabras clave

P. coatneyi; P. cynomolgi; P. inui; P. knowlesi LAMP; Primates; Zoonotic malaria

Texto completo

Añadir a Mi BVS

Imprimir

XML

PubMed Links

Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Plasmodium / ARN Ribosómico 18S / Zoonosis / Sensibilidad y Especificidad / Técnicas de Amplificación de Ácido Nucleico / Técnicas de Diagnóstico Molecular / Malaria Límite: Animals / Humans Idioma: En Revista: Acta Trop Año: 2024 Tipo del documento: Article País de afiliación: Malasia Pais de publicación: Países Bajos

Texto completo

Añadir a Mi BVS

Imprimir

XML

PubMed Links

Buscar en Google