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基于寡核苷酸分型和聚合酶链反应的菌株分型方法(肠杆菌基因间重复一致序列-聚合酶链反应、随机扩增多态性DNA-聚合酶链反应和外聚合酶链反应)

Spoligotyping and polymerase chain reaction based strains typing with methods (enterobacterial repetitive intergenic consensus-polymerase chain reaction, randomly amplified polymorphic dnas-polymerase chain reaction and out polymerase chain reaction).

作者信息

Sahin Funda, Tarhan G Lnur, Cinoglu Halil, Kayar Mediha Beg M, Yakici G Lfer

机构信息

Department of Medical Microbiology, Faculty of Medicine, Adiyaman University, Adiyaman, Turkey.

Tropical Diseases Research and Application Centeri, Çukurova University, Adana, Turkey.

出版信息

Int J Mycobacteriol. 2022 Jan-Mar;11(1):88-94. doi: 10.4103/ijmy.ijmy_253_21.

Abstract

BACKGROUND

In this study, it was aimed to investigate Mycobacterium bovis strains isolated from lungs and lymph nodes of slaughtered animals on clonal level by using different methods such as spoligotyping, enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR), randomly amplified polymorphic DNAs (RAPD-PCR) and OUT-PCR. Comparative evaluation of these methods was further conducted.

METHODS

A total of 38 M. bovis isolates were evaluated in the study. DNA isolation of all M. bovis strains isolated from pruvat free Löwenstein Jensen medium was done by boiling method for ERIC-PCR, RAPD-PCR, and OUT PCR. Mickle device was used for DNA isolation for spoligotyping method.

RESULTS

In 38 M. bovis isolates examined in our study, 4 different groups were determined by spoligotyping and RAPD-PCR test methods, and 5 different groups were detected in ERIC-PCR tests. In the OUT-PCR tests, the band which provides sufficient type separation was not observed.

CONCLUSION

ERIC-PCR, RAPD-PCR, and OUT-PCR methods are easily applicable, simple, and relatively inexpensive methods for evaluating the differences between origins in the typing of M. bovis. The tests need to be evaluated in more detail with extensive studies.

摘要

背景

本研究旨在通过使用不同方法,如间隔寡核苷酸分型(spoligotyping)、肠杆菌基因间重复共有序列聚合酶链反应(ERIC-PCR)、随机扩增多态性DNA(RAPD-PCR)和OUT-PCR,在克隆水平上研究从屠宰动物的肺和淋巴结中分离出的牛分枝杆菌菌株。并进一步对这些方法进行比较评估。

方法

本研究共评估了38株牛分枝杆菌分离株。从不含普鲁瓦特的罗-琴培养基中分离出的所有牛分枝杆菌菌株,采用煮沸法进行DNA提取,用于ERIC-PCR、RAPD-PCR和OUT-PCR。使用米克尔装置进行间隔寡核苷酸分型法的DNA提取。

结果

在我们研究中检测的38株牛分枝杆菌分离株中,通过间隔寡核苷酸分型和RAPD-PCR检测方法确定了4个不同的组,在ERIC-PCR检测中检测到5个不同的组。在OUT-PCR检测中,未观察到能提供足够分型分离的条带。

结论

ERIC-PCR、RAPD-PCR和OUT-PCR方法是评估牛分枝杆菌分型中不同来源差异的易于应用、简单且相对廉价的方法。这些检测需要通过广泛的研究进行更详细的评估。

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