A multiplex PCR assay for the differentiation of Mycobacterium tuberculosis complex reveals high rates of mixed-lineage tuberculosis infections among patients in Ghana.
A multiplex PCR assay for the differentiation of Mycobacterium tuberculosis complex reveals high rates of mixed-lineage tuberculosis infections among patients in Ghana.
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DOI：
10.3389/fcimb.2023.1125079
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发表时间：
2023
影响因子：
5.7
通讯作者：
中科院分区：
文献类型：
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在资源匮乏、结核病负担高的环境中，缺乏检测和鉴别结核分枝杆菌复体的快速诊断方法是影响结核病管理的一个主要挑战。本研究利用MTBC谱系的比较基因组分析；结核分枝杆菌，非洲分枝杆菌谱系5/6和牛分枝杆菌鉴定谱系特异性基因。引物设计用于开发多重PCR试验，该试验成功地区分了MTBC谱系。与其他呼吸道病原菌无交叉反应。使用临床样本对341名临床确诊的活动性结核病患者的痰DNA提取物进行了验证。其中结核分枝杆菌占24.9%，非洲分枝杆菌L5和L6分别占9.0%和14.4%。牛支原体感染是最不常见的谱系，为1.8%。另外，27.0%和17.0%的病例PCR阴性和未种。然而，混合系结核感染的记录却高达5.9%。这种多重PCR检测将允许在资源匮乏地区形成MTBC谱系，提供结核病感染的快速区分，以便在尽可能早的时间点选择适当的药物。它还将有助于流行病学监测研究，提供关于结核谱系流行情况的可靠信息，以及确定难以治疗的混合谱系结核感染病例。
In low-resource settings with high tuberculosis (TB) burdens, lack of rapid diagnostic methods for detection and differentiation of Mycobacterium tuberculosis complex (MTBC) is a major challenge affecting TB management. This study utilized comparative genomic analyses of MTBC lineages; M. tuberculosis, M. africanum Lineages 5/6 and M. bovis to identify lineage-specific genes. Primers were designed for the development of a Multiplex PCR assay which was successful in differentiating the MTBC lineages. There was no cross-reaction with other respiratory pathogens tested. Validation of the assay using clinical samples was performed with sputum DNA extracts from 341 clinically confirmed active TB patients. It was observed that 24.9% of cases were caused by M. tuberculosis, while M. africanum L5 & L6 reported 9.0% and 14.4%, respectively. M. bovis infection was the least frequently detected lineage with 1.8%. Also, 27.0% and 17.0% of the cases were PCR negative and unspeciated, respectively. However, mixed-lineage TB infections were recorded at a surprising 5.9%. This multiplex PCR assay will allow speciation of MTBC lineages in low-resource regions, providing rapid differentiation of TB infections to select appropriate medication at the earliest possible time point. It will also be useful in epidemiological surveillance studies providing reliable information on the prevalence of TB lineages as well as identifying difficult to treat cases of mixed-lineage tuberculosis infections.
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影响因子：
--
作者：
Agyare SA;Osei FA;Odoom SF;Mensah NK;Amanor E;Martyn-Dickens C;Owusu-Ansah M;Mohammed A;Yeboah EO
通讯作者：
Yeboah EO
影响因子：
--
作者：
de Jong, Bouke C.;Adetifa, Ifedayo;Adegbola, Richard A.
通讯作者：
Adegbola, Richard A.
影响因子：
9.4
作者：
de los Monteros, LEE;Galán, JC;Blázquez, J
通讯作者：
Blázquez, J
影响因子：
12.3
作者：
Brynildsrud O;Bohlin J;Scheffer L;Eldholm V
通讯作者：
Eldholm V
影响因子：
5.7
作者：
Akwani WC;van Vliet AHM;Joel JO;Andres S;Diricks M;Maurer FP;Chambers MA;Hingley-Wilson SM
通讯作者：
Hingley-Wilson SM