Increasing the Sensitivity of Leishmania RNA Virus 2 (LRV2) Detection with a Modification in cDNA Synthesis

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dc.contributor.authorKarakuş, Mehmet
dc.contributor.authorÖzbilgin, Ahmet
dc.contributor.authorTöz, Seray
dc.contributor.authorÖzbel, Yusuf
dc.contributor.authorNalçacı, Muhammed
dc.date.accessioned2023-01-12T20:32:16Z
dc.date.available2023-01-12T20:32:16Z
dc.date.issued2022
dc.departmentN/A/Departmenten_US
dc.description.abstractObjective: Leishmania RNA virus was detected the first time in the New World Leishmania species. Recent studies were also showed the presence of Leishmania RNA virus 2 (LRV2) in Old Word Leishmania species including Turkish L. major and L. tropica isolates. This study aimed to increase the sensitivity of qPCR with a modification in the denaturation step of cDNA preparation protocol. Methods: In this study, LRV2+ three L. major, two L. tropica strains and L. major control strain (MHOM/SU/73/5-ASKH) were included. Total RNA isolation was done using different numbers of Leishmania promastigotes (108 , 105 and 103 ). Before cDNA synthesis, samples were denatured at 95 °C for 2 min, as a modification of the kit procedure. qPCR was undertaken using 0.5 mM primers (LRV F-HR/LRV R-HR) diluted in SYBR Green Master mix. Results: We observed lower Ct values in amplicons with the modified version than with the classical kit protocol for cDNA synthesis, in all of the strains used in the study. The addition of pre-denaturation step at 95 °C showed lower Ct values meaning the sensitivity increased. Different parasite dilutions showed similar results. Conclusion: It is important to increase the sensitivity especially with the aim for detecting LRV in clinical samples obtained from patients probably have less number of parasites. The presence and burden of the virus can help to understand the relationship between the clinical findings and the pathogenicity of the parasite which may lead to changes in the course of treatment.en_US
dc.identifier.doi10.4274/tpd.galenos.2022.30074
dc.identifier.endpage90en_US
dc.identifier.issn2146-3077
dc.identifier.issue2en_US
dc.identifier.startpage86en_US
dc.identifier.trdizinid519179en_US
dc.identifier.urihttps://doi.org/10.4274/tpd.galenos.2022.30074
dc.identifier.urihttps://search.trdizin.gov.tr/yayin/detay/519179
dc.identifier.urihttps://hdl.handle.net/11454/81082
dc.identifier.volume46en_US
dc.indekslendigikaynakTR-Dizinen_US
dc.language.isoenen_US
dc.relation.ispartofTürkiye Parazitoloji Dergisien_US
dc.relation.publicationcategoryMakale - Ulusal Hakemli Dergi - Kurum Öğretim Elemanıen_US
dc.rightsinfo:eu-repo/semantics/openAccessen_US
dc.titleIncreasing the Sensitivity of Leishmania RNA Virus 2 (LRV2) Detection with a Modification in cDNA Synthesisen_US
dc.typeArticleen_US

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