TY  - JOUR
TI  - Increasing the Sensitivity of Leishmania RNA 
Virus 2 (LRV2) Detection with a Modification in 
cDNA Synthesis
AB  - Objective: 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.
AU  - Ozbilgin, Ahmet
AU  - Nalçacı, Muhammed
AU  - Töz, Seray
AU  - OZBEL, YUSUF
AU  - Karakus, Mehmet
DO  - 10.4274/tpd.galenos.2022.30074
PY  - 2022
JO  - Türkiye Parazitoloji Dergisi
VL  - 46
IS  - 2
SN  - 1300-6320
SP  - 86
EP  - 90
DB  - TRDizin
UR  - http://search/yayin/detay/519179
ER  -