[基耶省皮肤利什曼病患者分离的利什曼大利什曼原虫和热带利什曼原虫RNA病毒2的研究]。

IF 1.1 4区 医学 Q4 MICROBIOLOGY Mikrobiyoloji bulteni Pub Date : 2023-07-01 DOI:10.5578/mb.20239938
Cankut Karabulut, Tülay Aksoy, Ahmet Yıldırım, I Cüneyt Balcıoğlu
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引用次数: 0

摘要

利什曼RNA病毒(LRV)是一种双链RNA (dsRNA)病毒,被认为通过存在于利什曼原虫的许多分离株中,导致哺乳动物宿主中致病性利什曼原虫的严重炎症反应。在我们的研究中,旨在获得利什曼原虫RNA病毒2 (LRV2)存在的数据,该病毒被认为会导致利什曼病的临床过程发生变化。主要利什曼原虫和热带利什曼原虫分离自皮肤利什曼病(CL)患者在基耶省。在马尼萨Celal Bayar大学医学院寄生虫库冷冻液氮罐中保存的利什曼原虫菌株在合适的条件下复苏,并在NNN和rpm -1640培养基上培养。然后,将分离物置于26ºC培养箱中进入对数期,使用“高纯PCR模板制备试剂盒”进行DNA分离。利用利什曼原虫内部转录间隔-1 (ITS-1)基因区特异性引物和探针获得的dna,采用实时聚合酶链反应(Rt-PCR)熔融分析。从promastigote悬浮液中分离RNA后,通过逆转录合成cDNA。PCR扩增产物凝胶电泳后,以紫外光下LRV2阳性评价dsRNA条带形成情况。20株利什曼原虫分离株(10株热带利什曼原虫和10株大利什曼原虫)中,4株(3株热带利什曼原虫和1株大利什曼原虫)LRV2阳性。虽然在最近的研究中LRV的作用机制尚不完全清楚,但已知它加剧了疾病的临床,甚至对寄生虫耐药性的形成有影响。重要的是获得我国存在LRV的数据,并为将来的各种临床、药物开发、患病率研究、诊断和治疗该疾病做出贡献。
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[Investigation of Leishmania RNA Virus 2 in Leishmania major and Leishmania tropica Strains Isolated from Cutaneous Leishmaniasis Patients in Türkiye].

Leishmania RNA virus (LRV) is a double-stranded RNA (dsRNA) virus that is thought to contribute to the severe inflammatory response of the causative Leishmania parasite in the mammalian host by being present in many isolates of Leishmania spp. In our study, it was aimed to obtain data on the presence of Leishmania RNA Virus 2 (LRV2), which is thought to cause a change in the clinical course of leishmaniasis, in Leishmania major and Leishmania tropica isolates isolated from cutaneous leishmaniasis (CL) patients in Türkiye. Leishmania strains stored in liquid nitrogen tank by cryopreservation in Manisa Celal Bayar University Faculty of Medicine Parasite Bank were resuscitated under suitable conditions and cultivated in NNN and RPMI-1640 media. Then, the isolates were allowed to enter the logarithmic phase in a 26ºC incubator and DNA isolations were made using the "High Pure PCR Template Preparation Kit". Real-time polymerase chain reaction (Rt-PCR) melting analyzes were applied to the DNAs obtained by using primers and probes specific to the internal transcribed spacer-1 (ITS-1) gene region of Leishmania. After RNA isolation from promastigote suspension, cDNA synthesis was performed by reverse transcription. After gel electrophoresis with PCR amplification products, dsRNA band formation was evaluated in terms of LRV2 positivity under ultraviolet light. Among the 20 examined Leishmania spp. isolates (10 L.tropica and 10 L.major), four (three L.tropica, one L.major) were found to be positive for the presence of LRV2. Although the mechanism of LRV in recent studies has not been fully understood, it is known that it exacerbates the clinic of the disease and even has an effect on the formation of drug resistance by the parasite. It is important to obtain data on the presence of LRV in our country and to contribute to various clinical, drug development, prevalence studies, diagnosis and treatment of the disease in the future.

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来源期刊
Mikrobiyoloji bulteni
Mikrobiyoloji bulteni 生物-微生物学
CiteScore
1.60
自引率
20.00%
发文量
50
审稿时长
6-12 weeks
期刊介绍: Bulletin of Microbiology is the scientific official publication of Ankara Microbiology Society. It is published quarterly in January, April, July and October. The aim of Bulletin of Microbiology is to publish high quality scientific research articles on the subjects of medical and clinical microbiology. In addition, review articles, short communications and reports, case reports, editorials, letters to editor and other training-oriented scientific materials are also accepted. Publishing language is Turkish with a comprehensive English abstract. The editorial policy of the journal is based on independent, unbiased, and double-blinded peer-review. Specialists of medical and/or clinical microbiology, infectious disease and public health, and clinicians and researchers who are training and interesting with those subjects, are the target groups of Bulletin of Microbiology.
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