Chauhan Keerti, Gupta Rakesh Kumar, Anthwal Divya, Panwalkar Nikita, Desikan Prabha, Singhal Ritu, Bhalla Manpreet, Myneedu Vithal Prasad, Khayyam Khalid Umar, Shanmugam Siva Kumar, Chelvi K Silambu, Chandrasekaran Padmapriyadarsini, Giri Sidhartha, Turuk Jyotirmayee, Das Dasarathi, Pati Sanghamitra, Goyal Abhinav, Gupta Ashawant, Gupta Nalini Kant, Singh Manjula, Tyagi Jaya Sivaswami, Haldar Sagarika
Department of Experimental Medicine and Biotechnology, Postgraduate Institute of Medical Education and Research, Chandigarh, India.
Department of Microbiology, Bhopal Memorial Hospital and Research Centre, Bhopal, India.
Open Forum Infect Dis. 2025 Apr 11;12(7):ofaf207. doi: 10.1093/ofid/ofaf207. eCollection 2025 Jul.
We recently described the utility of the "TB Concentration & Transport" kit for biosafe, ambient temperature transport of dried sputum on -Filter, and the "TB DNA Extraction" kit for DNA extraction from -Filter for the early diagnosis of drug-resistant tuberculosis (TB). This study aimed to assess the feasibility and compatibility of these kits with line probe assay (LPA) under National Tuberculosis Elimination Programme (NTEP) settings.
Patients with presumptive pulmonary TB, multidrug-resistant (MDR) TB, or extensively drug-resistant TB (N = 8491) who attended Designated Microscopy Centers (DMCs, n = 13) under National Reference Laboratories (NRLs) at Bhopal, New Delhi, Chennai, and Bhubaneswar were screened by smear microscopy. The performance of -Filter extracted DNA-based LPA (Kit-LPA) was assessed against Direct-LPA on smear-positive sputum (n = 681), and feedback was obtained from scientists (n = 10) and laboratory technicians (n = 42) regarding logistics, kit usage, training, and troubleshooting. A scoring questionnaire was used to assess () the TB Concentration & Transport kit versus conventional sputum transport and () the TB DNA Extraction kit versus Hain GenoLyse kit (statistical significance of the scores was calculated using paired test).
Kit-LPA showed a sensitivity and specificity in the range of 89%-96% and 99%-100%, respectively, for rifampicin and isoniazid resistance detection and was comparable to Direct-LPA (concordance = 99%-100%; κ = 0.94-0.97). Overall scores indicated that () sputum transport on -Filters was more convenient as compared to conventional sputum transport ( < .0001) and () -Filter extracted DNA was easily amalgamated with LPA testing for MDR-TB detection.
These findings provide evidence for incorporation of these kits into the NTEP. Their use can facilitate sputum transport from DMCs to NRLs and provide universal drug susceptibility testing to people with TB residing in remote areas.
我们最近描述了“结核菌浓缩与转运”试剂盒用于在生物安全的环境温度下转运滤膜上的干燥痰液的效用，以及“结核菌DNA提取”试剂盒用于从滤膜上提取DNA以早期诊断耐多药结核病（TB）的效用。本研究旨在评估在国家结核病消除计划（NTEP）背景下这些试剂盒与线性探针分析（LPA）的可行性和兼容性。
在博帕尔、新德里、金奈和布巴内斯瓦尔的国家参考实验室（NRLs）下属的指定显微镜检查中心（DMCs，共13个）就诊的疑似肺结核、耐多药（MDR）结核病或广泛耐药结核病患者（N = 8491），通过涂片显微镜检查进行筛查。针对涂片阳性痰液（n = 681），评估基于滤膜提取DNA的LPA（试剂盒-LPA）相对于直接LPA的性能，并从科学家（n = 10）和实验室技术人员（n = 42）处获取关于物流、试剂盒使用、培训和故障排除的反馈。使用评分问卷来评估（1）结核菌浓缩与转运试剂盒与传统痰液转运的情况，以及（2）结核菌DNA提取试剂盒与海因基因裂解试剂盒的情况（使用配对t检验计算分数的统计学意义）。
试剂盒-LPA在检测利福平及异烟肼耐药性方面，敏感性和特异性分别在89% - 96%和99% - 100%范围内，与直接LPA相当（一致性 = 99% - 100%；κ = 0.94 - 0.97）。总体评分表明：（1）与传统痰液转运相比，滤膜上的痰液转运更方便（P <.0001）；（2）滤膜提取的DNA很容易与用于检测MDR - TB的LPA测试相结合。
这些发现为将这些试剂盒纳入NTEP提供了证据。它们的使用可以促进痰液从DMCs转运至NRLs，并为居住在偏远地区的结核病患者提供通用的药物敏感性检测。
