دورية أكاديمية
Increasing the Sensitivity of Leishmania RNA Virus 2 (LRV2) Detection with a Modification in cDNA Synthesis
| العنوان: | Increasing the Sensitivity of Leishmania RNA Virus 2 (LRV2) Detection with a Modification in cDNA Synthesis |
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| المؤلفون: | Muhammed Nalçacı, Mehmet Karakuş, Yusuf Özbel, Ahmet Özbilgin, Seray Töz |
| المصدر: | Türkiye Parazitoloji Dergisi, Vol 46, Iss 2, Pp 86-90 (2022) |
| بيانات النشر: | Galenos Yayinevi, 2022. |
| سنة النشر: | 2022 |
| المجموعة: | LCC:Medicine LCC:Infectious and parasitic diseases |
| مصطلحات موضوعية: | leishmania, leishmania rna virus 2, lrv2, cdna, qpcr, Medicine, Infectious and parasitic diseases, RC109-216 |
| الوصف: | 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. |
| نوع الوثيقة: | article |
| وصف الملف: | electronic resource |
| اللغة: | English Turkish |
| تدمد: | 1300-6320 2146-3077 |
| Relation: | http://www.turkiyeparazitolderg.org/archives/archive-detail/article-preview/ncreasing-the-sensitivity-of-ileishmania-i-rna-vir/51972; https://doaj.org/toc/1300-6320; https://doaj.org/toc/2146-3077 |
| DOI: | 10.4274/tpd.galenos.2022.30074 |
| URL الوصول: | https://doaj.org/article/82592f8c570a4c11aacb450496f05d5a |
| رقم الأكسشن: | edsdoj.82592f8c570a4c11aacb450496f05d5a |
| قاعدة البيانات: | Directory of Open Access Journals |
Full Text Finder | تدمد: | 13006320 21463077 |
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| DOI: | 10.4274/tpd.galenos.2022.30074 |