Abstract：
OBJECTIVE: Diagnosis of abdominal TB is an exigent task due to variable anatomical sites and non-specific clinical manifestations that closely resemble other diseases. Most of the available diagnostic modalities yield low sensitivities and need expertise to handle the specialized equipment. Hence, there is an urgent need to develop a rapid and reliable diagnostic test, so as to reduce the unnecessary morbidity. Therefore, we designed a multi-targeted loop-mediated isothermal amplification (MT-LAMP) for diagnosing abdominal TB.
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.
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.
摘要：
目的：腹部结核的诊断是一项紧迫的任务，因为不同的解剖部位和非特异性临床表现与其他疾病非常相似。大多数可用的诊断方式产生低灵敏度并且需要专业知识来处理专用设备。因此，迫切需要开发一种快速可靠的诊断测试，从而减少不必要的发病率。因此,我们设计了一种多靶向环介导等温扩增(MT-LAMP)诊断腹部结核的方法.
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。
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。
