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结核病实验室检测技术研究进展

17378 2020-03-12 12:08:08 王烁程,张林波 中国动物检疫
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结核病(tuberculosis,TB)是由结核分枝杆菌(Mycobacterium tuberculosis,MTB)引起的一种人兽共患慢性传染病,其早期诊断和检测非常困难。常规的TB实验室检测技术主要是痰涂片和痰培养,这是诊断肺结核的金标准,但均有一定的局限性,故对其优化和改良是目前TB检测研究的一个重要方向,目前主要集中在硬件设备的升级改造上。近年来,TB实验检测诊断技术发展较快。核酸扩增方法成为TB诊断研究的新方向,包括环介导等温扩增(loop-mediated isothermal amplification method,LAMP),结核分枝杆菌/利福平耐药实时荧光定量核酸扩增(Xpert Mycobacte-rium tuberculosis/Rifamfampin,Xpert MTB/RIF),交叉引物恒温扩增(crossing priming amplification,CPA),实时荧光恒温扩增(simultaneoous amplification and testing method,SAT)以及PCR技术等。免疫学检测技术是TB检测的另一个重要发展方向,现已研制出两种快速检测方法T-SPOT.TB和QuantiFERON-Gold test(QFT)。还有一些新技术正在研发中,如基质辅助激光解吸电离飞行时间质谱(matrix-assisted laser desorption/ionization time of flight mass spectrometry,MALDI-TOF-MS)以及代谢标记检测、纳米孔测序等。TB检测手段正由传统的细菌学检查转向免疫、分子诊断等更为精确的方法。然而,不同诊断方法均有局限性,在实际应用中需采用多种检测方法相结合的方式。本文对于全面了解多种TB检测技术的优点和局限性,从而选择合适的检测方法具有帮助意义。

Research Progress on Laboratory Technologies for Detection of Tuberculosis

Tuberculosis(TB)is a chronic zoonosis caused by mycobacterium tuberculosis(MTB),which is very difficult to diagnose and detect at its early stage. Fortunately,the technologies of TB laboratory testing and diagnosis has been developing rapidly in recent years. As the main conventional testing technologies,smear and cultivation of sputum are gold standards,but both of the two technologies have some limitations,hence,their optimization and improvement have become the important direction for TB testing,with an emphasis on upgrading of related hardware equipment. In order to solve such limitations,nucleic acid amplification methods have been studied and explored,including Loop-mediated Isothermal Amplification(LAMP),Xpert Mycobacterium tuberculosis/Rifampin(Xpert MTB/RIF),crossing priming amplification(CPA),simultaneous amplification and testing(SAT)and PCR. Immunological assays are also used for TB testing,in which,T-SPOT.TB and QuantiFERON-Gold test(QFT)have been already developed. Other new technologies are also in development,such as Matrix-assisted laser desorption ionization time-of-flight mass spectrometry(MALDI-TOF-MS),metabolic marker testing,nanopore sequencing,etc. At present,TB testing methods are turning to immunoassay,molecular diagnosis and other more accurate methods from traditional bacteriological testing. However,all the above methods are limited,therefore different technologies should be used together in practice. In conclusion,the advantages and limitations of various TB testing technologies were summarized,which would help to select appropriate method in practice.

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