{"title":"基于SYBR Green I反应的多靶点环介导等温扩增检测诊断胸膜结核:与GeneXpert®MTB/RIF检测的比较","authors":"Aishwarya Soni, Bhawna Dahiya, Abhishek Sheoran, Vipul Kumar, Astha Guliani, Nitin Kumar, Vikas Hooda, Aparna Yadav, Kiran Nehra, Promod K Mehta","doi":"10.1080/17476348.2023.2292738","DOIUrl":null,"url":null,"abstract":"<p><strong>Background: </strong>Diagnosis of pleural tuberculosis (TB) is tedious owing to its close resemblance with malignant pleural effusion and sparse bacterial load in clinical specimens. There is an immediate need to design a rapid and dependable diagnostic test to prevent unnecessary morbidity/mortality.</p><p><strong>Research design and methods: </strong>A multi-targeted loop-mediated isothermal amplification (MT-LAMP) was deliberated using mpt64 and IS6110 to diagnose pleural TB within pleural fluids/biopsies. MT-LAMP products were analyzed by gel-based and visual detection methods, viz. SYBR Green I, SYBR Green I+deoxyuridine triphosphate uracil-N-glycosylase (dUTP-UNG), and dry methyl green reactions.</p><p><strong>Results: </strong>In a pilot study, while assessing pleural TB/non-TB control subjects (<i>n</i> = 40), both SYBR Green I+dUTP-UNG/gel-based MT-LAMP assays exhibited better sensitivity/specificity than SYBR Green I and dry methyl green MT-LAMP. Since it is facile to work with SYBR Green I+dUTP-UNG than gel-based MT-LAMP, we validated the performance of SYBR Green I+dUTP-UNG in a higher number of specimens (<i>n</i> = 97), which revealed somewhat higher sensitivity (85.2 vs. 81.5%) and specificity (97.7 vs. 90.7%) than SYBR Green I MT-LAMP. Furthermore, the sensitivity attained by SYBR Green I+dUTP-UNG MT-LAMP was significantly higher (<i>p</i> < 0.001) than GeneXpert.</p><p><strong>Conclusions: </strong>Our SYBR Green I+dUTP-UNG MT-LAMP is a simple and reliable method to diagnose pleural TB, which may translate into a point-of-care test.</p>","PeriodicalId":94007,"journal":{"name":"Expert review of respiratory medicine","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"2023-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Diagnosis of pleural tuberculosis by multi-targeted loop-mediated isothermal amplification assay based on SYBR Green I reaction: comparison with GeneXpert® MTB/RIF assay.\",\"authors\":\"Aishwarya Soni, Bhawna Dahiya, Abhishek Sheoran, Vipul Kumar, Astha Guliani, Nitin Kumar, Vikas Hooda, Aparna Yadav, Kiran Nehra, Promod K Mehta\",\"doi\":\"10.1080/17476348.2023.2292738\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Background: </strong>Diagnosis of pleural tuberculosis (TB) is tedious owing to its close resemblance with malignant pleural effusion and sparse bacterial load in clinical specimens. There is an immediate need to design a rapid and dependable diagnostic test to prevent unnecessary morbidity/mortality.</p><p><strong>Research design and methods: </strong>A multi-targeted loop-mediated isothermal amplification (MT-LAMP) was deliberated using mpt64 and IS6110 to diagnose pleural TB within pleural fluids/biopsies. MT-LAMP products were analyzed by gel-based and visual detection methods, viz. SYBR Green I, SYBR Green I+deoxyuridine triphosphate uracil-N-glycosylase (dUTP-UNG), and dry methyl green reactions.</p><p><strong>Results: </strong>In a pilot study, while assessing pleural TB/non-TB control subjects (<i>n</i> = 40), both SYBR Green I+dUTP-UNG/gel-based MT-LAMP assays exhibited better sensitivity/specificity than SYBR Green I and dry methyl green MT-LAMP. Since it is facile to work with SYBR Green I+dUTP-UNG than gel-based MT-LAMP, we validated the performance of SYBR Green I+dUTP-UNG in a higher number of specimens (<i>n</i> = 97), which revealed somewhat higher sensitivity (85.2 vs. 81.5%) and specificity (97.7 vs. 90.7%) than SYBR Green I MT-LAMP. Furthermore, the sensitivity attained by SYBR Green I+dUTP-UNG MT-LAMP was significantly higher (<i>p</i> < 0.001) than GeneXpert.</p><p><strong>Conclusions: </strong>Our SYBR Green I+dUTP-UNG MT-LAMP is a simple and reliable method to diagnose pleural TB, which may translate into a point-of-care test.</p>\",\"PeriodicalId\":94007,\"journal\":{\"name\":\"Expert review of respiratory medicine\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2023-07-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Expert review of respiratory medicine\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.1080/17476348.2023.2292738\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2023/12/26 0:00:00\",\"PubModel\":\"Epub\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Expert review of respiratory medicine","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1080/17476348.2023.2292738","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2023/12/26 0:00:00","PubModel":"Epub","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0
摘要
背景:胸膜结核(TB)的诊断非常繁琐，因为它与恶性胸腔积液非常相似，而且临床标本中细菌负荷稀少。迫切需要设计一种快速可靠的诊断测试，以防止不必要的发病/死亡。研究设计和方法:采用mpt64和IS6110进行多靶点环介导等温扩增(MT-LAMP)诊断胸膜结核/活检。MT-LAMP产品通过凝胶和视觉检测方法，即SYBR Green I、SYBR Green I+脱氧尿苷三磷酸尿嘧啶n -糖基化酶(dUTP-UNG)和干甲基绿反应进行分析。结果:在一项初步研究中，在评估胸膜结核/非结核对照受试者(n = 40)时，SYBR Green I+dUTP-UNG/凝胶基MT-LAMP检测比SYBR Green I和干甲基绿MT-LAMP表现出更好的敏感性/特异性。由于SYBR Green+dUTP-UNG比基于凝胶的MT-LAMP更容易工作，我们在更多的标本(n = 97)中验证了SYBR Green+dUTP-UNG的性能，结果显示，与SYBR Green MT-LAMP相比，SYBR Green+dUTP-UNG的灵敏度(85.2比81.5%)和特异性(97.7比90.7%)有所提高。结论:SYBR Green I+dUTP-UNG MT-LAMP是一种简单可靠的胸膜结核诊断方法，可以转化为一种即时检测方法。
Diagnosis of pleural tuberculosis by multi-targeted loop-mediated isothermal amplification assay based on SYBR Green I reaction: comparison with GeneXpert® MTB/RIF assay.
Background: Diagnosis of pleural tuberculosis (TB) is tedious owing to its close resemblance with malignant pleural effusion and sparse bacterial load in clinical specimens. There is an immediate need to design a rapid and dependable diagnostic test to prevent unnecessary morbidity/mortality.
Research design and methods: A multi-targeted loop-mediated isothermal amplification (MT-LAMP) was deliberated using mpt64 and IS6110 to diagnose pleural TB within pleural fluids/biopsies. MT-LAMP products were analyzed by gel-based and visual detection methods, viz. SYBR Green I, SYBR Green I+deoxyuridine triphosphate uracil-N-glycosylase (dUTP-UNG), and dry methyl green reactions.
Results: In a pilot study, while assessing pleural TB/non-TB control subjects (n = 40), both SYBR Green I+dUTP-UNG/gel-based MT-LAMP assays exhibited better sensitivity/specificity than SYBR Green I and dry methyl green MT-LAMP. Since it is facile to work with SYBR Green I+dUTP-UNG than gel-based MT-LAMP, we validated the performance of SYBR Green I+dUTP-UNG in a higher number of specimens (n = 97), which revealed somewhat higher sensitivity (85.2 vs. 81.5%) and specificity (97.7 vs. 90.7%) than SYBR Green I MT-LAMP. Furthermore, the sensitivity attained by SYBR Green I+dUTP-UNG MT-LAMP was significantly higher (p < 0.001) than GeneXpert.
Conclusions: Our SYBR Green I+dUTP-UNG MT-LAMP is a simple and reliable method to diagnose pleural TB, which may translate into a point-of-care test.
求助内容：
应助结果提醒方式：
京公网安备 11010802042870号
