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

Ozbilgin, Ahmet; Nalçacı, Muhammed; Töz, Seray; OZBEL, YUSUF; Karakus, Mehmet

doi:10.4274/tpd.galenos.2022.30074

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

Muhammed NALÇACI, (Ege Üniversitesi, Fen Bilimleri Enstitüsü, Biyoloji Bölümü, İzmir, Türkiye)

Mehmet KARAKUŞ, (Türkiye Sağlık Bilimleri Üniversitesi, Hamidiye Tıp Fakültesi, Tıbbi Mikrobiyoloji Anabilim Dalı, İstanbul, Türkiye)

Yusuf ÖZBEL, (Ege Üniversitesi, Tıp Fakültesi, Parazitoloji Anabilim Dalı, İzmir, Türkiye)

Ahmet ÖZBİLGİN, (Celal Bayar Üniversitesi, Tıp Fakültesi, Parazitoloji Anabilim Dalı, Manisa, Türkiye)

Seray TÖZ (Ege Üniversitesi, Tıp Fakültesi, Parazitoloji Anabilim Dalı, İzmir, Türkiye)

Türkiye Parazitoloji Dergisi

2 1

Yıl: 2022 Cilt: 46 Sayı: 2 Sayfa Aralığı: 86 - 90 Metin Dili: İngilizce DOI: 10.4274/tpd.galenos.2022.30074 İndeks Tarihi: 15-06-2022

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

Öz:

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.

Anahtar Kelime:

Leishmania RNA Virüs 2 (LRV2) Saptanmasında cDNA Sentezindeki BirModifikasyon ile Hassasiyetin Artırılması

Öz:

Sağlık Bilimleri Üniversitesi Türkiye Hamidiye Tıp Fakültesi Tıbbi Mikrobiyoloji Anabilim Dalı, İstanbul, Türkiye

Anahtar Kelime:

Belge Türü: Makale Makale Türü: Araştırma Makalesi Erişim Türü: Erişime Açık

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APA	Nalçacı M, Karakus M, OZBEL Y, Ozbilgin A, Töz S (2022). Increasing the Sensitivity of Leishmania RNA Virus 2 (LRV2) Detection with a Modification in cDNA Synthesis. , 86 - 90. 10.4274/tpd.galenos.2022.30074
Chicago	Nalçacı Muhammed,Karakus Mehmet,OZBEL YUSUF,Ozbilgin Ahmet,Töz Seray Increasing the Sensitivity of Leishmania RNA Virus 2 (LRV2) Detection with a Modification in cDNA Synthesis. (2022): 86 - 90. 10.4274/tpd.galenos.2022.30074
MLA	Nalçacı Muhammed,Karakus Mehmet,OZBEL YUSUF,Ozbilgin Ahmet,Töz Seray Increasing the Sensitivity of Leishmania RNA Virus 2 (LRV2) Detection with a Modification in cDNA Synthesis. , 2022, ss.86 - 90. 10.4274/tpd.galenos.2022.30074
AMA	Nalçacı M,Karakus M,OZBEL Y,Ozbilgin A,Töz S Increasing the Sensitivity of Leishmania RNA Virus 2 (LRV2) Detection with a Modification in cDNA Synthesis. . 2022; 86 - 90. 10.4274/tpd.galenos.2022.30074
Vancouver	Nalçacı M,Karakus M,OZBEL Y,Ozbilgin A,Töz S Increasing the Sensitivity of Leishmania RNA Virus 2 (LRV2) Detection with a Modification in cDNA Synthesis. . 2022; 86 - 90. 10.4274/tpd.galenos.2022.30074
IEEE	Nalçacı M,Karakus M,OZBEL Y,Ozbilgin A,Töz S "Increasing the Sensitivity of Leishmania RNA Virus 2 (LRV2) Detection with a Modification in cDNA Synthesis." , ss.86 - 90, 2022. 10.4274/tpd.galenos.2022.30074
ISNAD	Nalçacı, Muhammed vd. "Increasing the Sensitivity of Leishmania RNA Virus 2 (LRV2) Detection with a Modification in cDNA Synthesis". (2022), 86-90. https://doi.org/10.4274/tpd.galenos.2022.30074

APA	Nalçacı M, Karakus M, OZBEL Y, Ozbilgin A, Töz S (2022). Increasing the Sensitivity of Leishmania RNA Virus 2 (LRV2) Detection with a Modification in cDNA Synthesis. Türkiye Parazitoloji Dergisi, 46(2), 86 - 90. 10.4274/tpd.galenos.2022.30074
Chicago	Nalçacı Muhammed,Karakus Mehmet,OZBEL YUSUF,Ozbilgin Ahmet,Töz Seray Increasing the Sensitivity of Leishmania RNA Virus 2 (LRV2) Detection with a Modification in cDNA Synthesis. Türkiye Parazitoloji Dergisi 46, no.2 (2022): 86 - 90. 10.4274/tpd.galenos.2022.30074
MLA	Nalçacı Muhammed,Karakus Mehmet,OZBEL YUSUF,Ozbilgin Ahmet,Töz Seray Increasing the Sensitivity of Leishmania RNA Virus 2 (LRV2) Detection with a Modification in cDNA Synthesis. Türkiye Parazitoloji Dergisi, vol.46, no.2, 2022, ss.86 - 90. 10.4274/tpd.galenos.2022.30074
AMA	Nalçacı M,Karakus M,OZBEL Y,Ozbilgin A,Töz S Increasing the Sensitivity of Leishmania RNA Virus 2 (LRV2) Detection with a Modification in cDNA Synthesis. Türkiye Parazitoloji Dergisi. 2022; 46(2): 86 - 90. 10.4274/tpd.galenos.2022.30074
Vancouver	Nalçacı M,Karakus M,OZBEL Y,Ozbilgin A,Töz S Increasing the Sensitivity of Leishmania RNA Virus 2 (LRV2) Detection with a Modification in cDNA Synthesis. Türkiye Parazitoloji Dergisi. 2022; 46(2): 86 - 90. 10.4274/tpd.galenos.2022.30074
IEEE	Nalçacı M,Karakus M,OZBEL Y,Ozbilgin A,Töz S "Increasing the Sensitivity of Leishmania RNA Virus 2 (LRV2) Detection with a Modification in cDNA Synthesis." Türkiye Parazitoloji Dergisi, 46, ss.86 - 90, 2022. 10.4274/tpd.galenos.2022.30074
ISNAD	Nalçacı, Muhammed vd. "Increasing the Sensitivity of Leishmania RNA Virus 2 (LRV2) Detection with a Modification in cDNA Synthesis". Türkiye Parazitoloji Dergisi 46/2 (2022), 86-90. https://doi.org/10.4274/tpd.galenos.2022.30074