METHODS: We evaluated an MT-LAMP (using mpt64 and IS6110) to diagnose abdominal TB within ascitic fluids and intestinal/peritoneal biopsies and compared these results with multiplex-PCR (M-PCR) using the same targets. MT-LAMP products were analyzed by gel electrophoresis and visual detection methods, that is, hydroxy naphthol blue and SYBR Green I reaction.
RESULTS: Sensitivities of 80.9% and 84.6% were obtained in suspected (n = 42) and total abdominal TB (n = 52) cases, respectively by gel-based MT-LAMP, with 97.3% (n = 37) specificity in non-TB controls. Notably, sensitivities attained by gel-based/SYBR Green I MT-LAMP in both clinically suspected and total abdominal TB cases were significantly higher (P < 0.05) than M-PCR. Furthermore, sensitivity obtained with SYBR Green I was equivalent to that of gel-based MT-LAMP, while somewhat lesser specificity (94.6%) was attained with SYBR Green I, compared with gel-based MT-LAMP.
CONCLUSIONS: Both gel-based and SYBR Green MT-LAMP exhibited equivalent sensitivities to diagnose abdominal TB. Because SYBR Green LAMP is easier to perform than a gel-based assay, we are currently focused on improving the specificity of this assay so as to develop a diagnostic kit.
方法:我们评估了MT-LAMP(使用mpt64和IS6110)在腹水和肠/腹膜活检中诊断腹部结核,并将这些结果与使用相同靶标的多重PCR(M-PCR)进行比较。采用凝胶电泳和视觉检测方法对MT-LAMP产品进行分析,也就是说,羟基萘酚蓝与SYBR绿Ⅰ反应。
结果:在疑似(n=42)和全腹结核(n=52)病例中,敏感性分别为80.9%和84.6%。分别通过基于凝胶的MT-LAMP,在非TB对照中具有97.3%(n=37)的特异性。值得注意的是,基于凝胶/SYBRGreenIMT-LAMP在临床可疑和全腹TB病例中的敏感性均显着高于M-PCR(P<0.05)。此外,用SYBRGreenI获得的灵敏度与基于凝胶的MT-LAMP相当,而SYBRGreenI的特异性稍低(94.6%),与基于凝胶的MT-LAMP相比。
结论:基于凝胶的和SYBRGreenMT-LAMP对诊断腹部TB表现出同等的敏感性。因为SYBRGreenLAMP比基于凝胶的检测更容易执行,我们目前专注于提高该测定的特异性,以开发诊断试剂盒。