| Journal: | Memorias do Instituto Oswaldo Cruz |
| Database: | PERIÓDICA |
| System number: | 000400860 |
| ISSN: | 0074-0276 |
| Authors: | Tiwananthagorn, Saruda1 Kato, Hirotomo2 Yeewa, Ranchana1 Muengpan, Amontip1 Polseela, Raxsina3 Leelayoova, Saovanee4 |
| Institutions: | 1Chiang Mai University, Faculty of Veterinary Medicine, Muang, Chiang Mai. Tailandia 2Jichi Medical University, Division of Medical Zoology, Tochigi. Japón 3Naresuan University, Faculty of Medical Science, Phitsanulok. Tailandia 4Phramongkutklao College of Medicine, Department of Parasitology, Bangkok, Phra Nakhon. Tailandia |
| Year: | 2017 |
| Season: | Feb |
| Volumen: | 112 |
| Number: | 2 |
| Pages: | 100-107 |
| Country: | Brasil |
| Language: | Inglés |
| Document type: | Artículo |
| Approach: | Experimental, aplicado |
| English abstract | Leishmaniasis caused by Leishmania martiniquensis infection has been reported in human and domestic animals of Martinique Island, Germany, Switzerland, USA, Myanmar and Thailand. The peculiar clinical features of disseminated cutaneous and visceral forms co-existence render the urgent need of specific diagnostic tool to identify the natural sand fly vectors for effective prevention and control strategies. Loop-mediated isothermal amplification (LAMP) of 18S rRNA gene as well as polymerase chain reaction (PCR) of minicircle kinetoplast DNA gene (PCR-mkDNA) have never been applied to detect L. martiniquensis and L. siamensis in sand fly vectors. OBJECTIVE The present study was aimed to validate malachite green-LAMP (MG-LAMP) and PCR-mkDNA techniques to detect L. martiniquensis in sand fly vectors, compared with the conventional PCR of internal transcribed spacer 1 (PCR-ITS1). METHODS We compared the validity of LAMP of 18S rRNA gene and PCR-mkDNA, to PCR-ITS1 in simulation model of L. martiniquensis infection in Sergentomyia gemmea sand flies. Attributable to the sensitivity and specificity, PCR-mkDNA was consecutively applied to detect L. martiniquensis in 380 female sand fly individuals captured in the newly identified affected region of Lamphun Province, Thailand. FINDINGS AND MAIN CONCLUSIONS Results showed that PCR-mkDNA could detect at least one promastigote per sand fly, which was 10-time superior to LAMP and PCR-ITS1. In addition, PCR-mkDNA was more specific, able to differentiate L. martiniquensis from other viscerotropic Leishmania species, such as L. siamensis, L. (L.) donovani, and L. (L.) infantum. Consecutively, mass screening of L. martiniquensis in 380 female sand fly individuals by PCR-mkDNA was implemented in a new affected area of Thailand where a patient with leishmaniasis/HIV co-infection resides; however Leishmania DNA was undetected. In conclusion, PCR-mkDNA is a promising tool for molecular mass screening |
| Disciplines: | Medicina |
| Keyword: | Microbiología, Insectos, Leishmaniasis, Vectores biológicos, Monitoreo molecular, Reacción en cadena de la polimerasa (PCR), Amplificación génica, Cinetoplástidos, ADN, Leishmania martiniquensis |
| Keyword: | Medicine, Microbiology, Insects, Leishmaniasis, Biological vectors, Molecular screening, Polymerase chain reaction (PCR), Gene amplification, Kinetoplastids, DNA, Leishmania martiniquensis |
| Full text: | Texto completo (Ver HTML) |
