| Autor: |
Kim J. M. van Bergen, Eric A.T. Brienen, Bodo S. Randrianasolo, Charles E. Ramarokoto, Peter Leutscher, Eyrun F. Kjetland, Angela van Diepen, Floris Dekker, Vittorio Saggiomo, Aldrik H. Velders, Lisette van Lieshout |
| Jazyk: |
angličtina |
| Rok vydání: |
2024 |
| Předmět: |
|
| Zdroj: |
Frontiers in Parasitology, Vol 3 (2024) |
| Druh dokumentu: |
article |
| ISSN: |
2813-2424 |
| DOI: |
10.3389/fpara.2024.1297310 |
| Popis: |
Detection of Schistosoma spp. DNA in gynaecological samples by quantitative real-time polymerase chain reaction (qPCR) is considered to be the reference diagnostic test for female genital schistosomiasis (FGS). However, qPCR needs expensive laboratory procedures and highly trained technicians. Loop-mediated amplification (LAMP) is a more field-friendly isothermal procedure for the detection of parasite-specific DNA, but it still requires electrically powered equipment. Here, we validated a Schistosoma haematobium-specific Sh-LAMP procedure and tested a fully instrument-free isothermal amplification using a novel low-cost, and reusable Temperature-cup (T-cup) device. Specific primers were selected based on published assays, targeting the ribosomal intergenic spacer (IGS) region of S. haematobium. Technical validation of the IGS-Sh-LAMP was performed using 20 negative controls, including DNA extracts of soil-transmitted helminths and S. mansoni, and a 10-fold dilution series (100–10−3) of DNA extracted from a single S. haematobium egg (n=4). For clinical validation, the IGS-Sh-LAMP was tested on 125 DNA samples extracted from vaginal swabs of a previous FGS study in Madagascar. Results were compared with the quantification cycle value (Cq) of the standard ITS-2 targeting qPCR. Single S. haematobium egg DNA up to a 10–2 dilution and an ITS-2 Cq |
| Databáze: |
Directory of Open Access Journals |
| Externí odkaz: |
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