ArticleParasites & vectors2020
Evaluation of a pan-Leishmania SL RNA qPCR assay for parasite detection in laboratory-reared and field-collected sand flies and reservoir hosts.
Article in Parasites & vectors, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers, 2 of them syntheses that pooled it.
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12 citing papers in PubMed, 2 syntheses or guidelines pooled it, 17 citations in OpenAlex.
- Performance of recombinase polymerase amplification assay for cutaneous leishmaniasis detection: a systematic review and meta-analysis.BMC infectious diseases · 2026Pooled it
- World Association for Veterinary Dermatology Consensus Statement for Diagnosis, and Evidence-Based Clinical Practice Guidelines for Treatment and Prevention of Canine Leishmaniosis.Veterinary dermatology · 2025Guideline
- Integrating molecular tools into leishmaniosis surveillance: evaluation of a commercial qPCR kit in sand fly vectors.Parasites & vectors · 2026Article
- From Skin to Sequence: Pan-species Detection and Speciation of Leishmania.The Journal of infectious diseases · 2026Article
- Molecular survey and risk factors ofParasitology · 2026Article
- Pilot study to assess the impact of feed-through insecticide on the sand fly density in an endemic area of zoonotic cutaneous leishmaniasis in Morocco.PLoS neglected tropical diseases · 2025Article
- A one-step multiplex qPCR assay for simultaneous identification and quantification of Leishmania martiniquensis and Leishmania orientalis/Leishmania chancei and detection and quantification of trypanosomatids in clinical samples.Parasite (Paris, France) · 2025Article
- Evaluation of Loopamp Leishmania detection kit for the diagnosis of cutaneous leishmaniasis in Ethiopia.Parasites & vectors · 2024Article
- PCR for detection of Leishmania donovani from microscopically negative tissue smears of suspected patients in Gondar, Ethiopia.PLoS neglected tropical diseases · 2023Article
- Concurrent molecular characterization of sand flies and Leishmania parasites by amplicon-based next-generation sequencing.Parasites & vectors · 2022Article
- Evaluation of molecular assays to detect Leishmania donovani in Phlebotomus argentipes fed on post-kala-azar dermal leishmaniasis patients.Parasites & vectors · 2021Article
- Evaluation of a kDNA-Based qPCR Assay for the Detection and Quantification of Old WorldMicroorganisms · 2020Article
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Authors and funding
10 authors at 2 institutions in 2 countries.
Funding
Abstract
backgroundIn eco-epidemiological studies, Leishmania detection in vectors and reservoirs is frequently accomplished by high-throughput and sensitive molecular methods that target minicircle kinetoplast DNA (kDNA). A pan-Leishmania SYBR green quantitative PCR (qPCR) assay which detects the conserved spliced-leader RNA (SL RNA) sequence was developed recently. This study assessed the SL RNA assay performance combined with a crude extraction method for the detection of Leishmania in field-collected and laboratory-reared sand flies and in tissue samples from hyraxes as reservoir hosts.
methodsField-collected and laboratory-infected sand fly and hyrax extracts were subjected to three different qPCR approaches to assess the suitability of the SL RNA target for Leishmania detection. Nucleic acids of experimentally infected sand flies were isolated with a crude extraction buffer with ethanol precipitation and a commercial kit and tested for downstream DNA and RNA detection. Promastigotes were isolated from culture and sand fly midguts to assess whether there was difference in SL RNA and kDNA copy numbers. Naive sand flies were spiked with a serial dilution of promastigotes to make a standard curve.
resultsThe qPCR targeting SL RNA performed well on infected sand fly samples, despite preservation and extraction under presumed unfavorable conditions for downstream RNA detection. Nucleic acid extraction by a crude extraction buffer combined with a precipitation step was highly compatible with downstream SL RNA and kDNA detection. Copy numbers of kDNA were found to be identical in culture-derived parasites and promastigotes isolated from sand fly midguts. SL RNA levels were slightly lower in sand fly promastigotes (ΔCq 1.7). The theoretical limit of detection and quantification of the SL RNA qPCR respectively reached down to 10
conclusionsThis study shows that a crude extraction method in combination with the SL RNA qPCR assay is suitable for the detection and quantification of Leishmania in sand flies. The assay is inexpensive, sensitive and pan-Leishmania specific, and accordingly an excellent assay for high-throughput screening in entomological research.
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