New mini-satellite tandem repeat markers for strain typing of Mycobacterium avium subsp. paratuberculosis.

IF 2.8 4区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Molecular Biology Reports Pub Date : 2025-07-14 DOI:10.1007/s11033-025-10792-w

Elika Faraj Tabrizi, Mohammad Reza Zolfaghari, Nader Mosavari, Razieh Nazari, Keyvan Tadayon

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引用次数: 0

Abstract

Background: The multiplicity of infection sources along with the genetically homogenous nature of Mycobacterium avium subsp. paratuberculosis (MAP) necessitates the application of high-discriminative fingerprinting techniques to establish epidemiological links and control of paratuberculosis in ruminants. The present study aimed to develop a simple Multiple-Locus Variable-Number Tandem-Repeat Analysis (MLVA) assay suitable for low-budget laboratory settings.

Methods: A bioinformatics screening was conducted on three annotated MAP genomes (K10, MAP4, and MAP-JIII-386) leading to the detection of 7 potential polymorphic variable-number tandem repeats (VNTRs). A MAP collection of the three aforementioned strains, 30 field isolates and 2 laboratory strains from Iran plus 59 global Mycobacterium avium complex strains were subsequently genotyped at laboratory or in situ. Findings were validated against the French INRAE MLVA typing system (Thibault's scheme) dedicated to typing Mycobacterium avium subsp. paratuberculosis, hominissuis, avium, and silvaticum.

Results: Fourty-one MLVA types were detected by our suggested MLVA scheme, compared to 45 types found by Thibault's scheme. The calculated discriminatory power of the MLVA for the former and the latter was 0.93 and 0.95, respectively.

Conclusion: The suggested MLVA scheme discriminated between isolates with identical types determined by Thibault's scheme. Noting the fact that strains grouped in INMV types 2, 3, 4, 5, 33, 39, 68, 82, 131, 219, 220, New1 and New 25 were further resolved into sub-groups, we suggest considering VNTR 1067 (DI = 0.71), MAP 4356 (DI = 0.68), MATR 7 (DI = 0.53), MAP 3824 (DI = 0.56) and MATR 6 (DI = 0.49), in any future upgrading of the current version of the Thibault's scheme.

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禽分枝杆菌亚种分型的新型微型卫星串联重复序列标记。副结核。

背景：感染源的多样性以及禽分枝杆菌亚种的遗传同质性。副结核（MAP）需要应用高鉴别指纹技术来建立反刍动物副结核的流行病学联系和控制。本研究旨在开发一种简单的多位点可变数串联重复分析（MLVA）方法，适用于低预算的实验室环境。方法：对3个带注释的MAP基因组（K10、MAP4和MAP- jiii -386）进行生物信息学筛选，检测出7个潜在多态性可变数串联重复序列（VNTRs）。MAP收集了上述3株菌株、30株现场分离株和2株来自伊朗的实验室菌株以及59株全球禽分枝杆菌复合体菌株，随后在实验室或原位进行了基因分型。研究结果通过法国INRAE MLVA分型系统（Thibault方案）进行了验证，该系统专门用于分型鸟分枝杆菌亚种。副结核菌病，人痘病，鸟病和银屑病。结果：我们建议的MLVA方案检测到41种MLVA类型，而Thibault方案检测到45种。前者和后者的MLVA的计算区分力分别为0.93和0.95。结论：所建议的MLVA方案可区分由Thibault方案确定的相同类型的分离株。考虑到INMV类型2、3、4、5、33、39、68、82、131、219、220、New1和New 25被进一步细分为亚群，我们建议在未来升级当前版本的Thibault’s方案时，考虑VNTR 1067 （DI = 0.71）、MAP 4356 （DI = 0.68）、MATR 7 （DI = 0.53）、MAP 3824 （DI = 0.56）和MATR 6 （DI = 0.49）。

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来源期刊

Molecular Biology Reports 生物-生化与分子生物学

CiteScore

5.00

自引率

0.00%

发文量

1048

审稿时长

5.6 months

期刊介绍： Molecular Biology Reports publishes original research papers and review articles that demonstrate novel molecular and cellular findings in both eukaryotes (animals, plants, algae, funghi) and prokaryotes (bacteria and archaea).The journal publishes results of both fundamental and translational research as well as new techniques that advance experimental progress in the field and presents original research papers, short communications and (mini-) reviews.