A new droplet digital PCR assay: improving detection of paucibacillary smear-negative pulmonary tuberculosis
Objectives: Smear-negative pulmonary TB (PTB) is difficult to diagnose. Current diagnosis and treatment monitoring methods have inherent limitations. Droplet digital PCR (ddPCR) is a new technique with high sensitivity. This study presents a novel ddPCR for rapid and sensitive identification of Myco...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2022-09-01
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| Series: | International Journal of Infectious Diseases |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S1201971222004374 |
| _version_ | 1811297384916844544 |
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| author | Zhenzhen Zhao Tao Wu Minjin Wang Xiaojuan Chen Tangyuheng Liu Yanjun Si Yanhong Zhou Binwu Ying |
| author_facet | Zhenzhen Zhao Tao Wu Minjin Wang Xiaojuan Chen Tangyuheng Liu Yanjun Si Yanhong Zhou Binwu Ying |
| author_sort | Zhenzhen Zhao |
| collection | DOAJ |
| description | Objectives: Smear-negative pulmonary TB (PTB) is difficult to diagnose. Current diagnosis and treatment monitoring methods have inherent limitations. Droplet digital PCR (ddPCR) is a new technique with high sensitivity. This study presents a novel ddPCR for rapid and sensitive identification of Mycobacterium tuberculosis (MTB). Methods: MTB DNA was detected in respiratory specimens from suspected PTB cases using ddPCR assay, which was directed at two different locations within IS6110. We, for the first time, evaluated the clinical diagnostic ability of this ddPCR for paucibacillary smear-negative PTB. Results: A total of 605 PTB suspects were recruited, including 263 patients with confirmed PTB (84.03% from smear-negative PTB) and 342 without PTB. The sensitivity and specificity of IS6110 ddPCR were 61.22% (95% confidence interval (CI) 55.00-67.10%) and 95.03% (95% CI 92.20-97.10%) for total PTB and 57.92% (95% CI 51.10-64.50%) and 94.57% (95% CI 91.20-96.90%) for smear-negative PTB. ddPCR assay outperformed Xpert MTB/RIF (53.08% vs 28.46%, P = 0.020) in smear-negative PTB detection. Furthermore, effective anti-TB treatment was linked to significantly lower IS6110 copies detected by ddPCR. Conclusion: Herein, we developed and validated a highly sensitive and robust ddPCR assay for MTB quantification in respiratory specimens, which improves diagnosis and therapeutic effect evaluation of smear-negative PTB. |
| first_indexed | 2024-04-13T06:03:25Z |
| format | Article |
| id | doaj.art-9d984a53b22b417c867616715d7a436a |
| institution | Directory Open Access Journal |
| issn | 1201-9712 |
| language | English |
| last_indexed | 2024-04-13T06:03:25Z |
| publishDate | 2022-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | International Journal of Infectious Diseases |
| spelling | doaj.art-9d984a53b22b417c867616715d7a436a2022-12-22T02:59:20ZengElsevierInternational Journal of Infectious Diseases1201-97122022-09-01122820828A new droplet digital PCR assay: improving detection of paucibacillary smear-negative pulmonary tuberculosisZhenzhen Zhao0Tao Wu1Minjin Wang2Xiaojuan Chen3Tangyuheng Liu4Yanjun Si5Yanhong Zhou6Binwu Ying7Department of Laboratory Medicine, West China Hospital, Sichuan University, Chengdu, Sichuan, 610041, ChinaDepartment of Clinical Laboratory Medicine, People's Hospital of Ningxia Hui Autonomous Region (First Affiliated Hospital of Northwest Minzu University), Yinchuan, 750002, ChinaDepartment of Laboratory Medicine, West China Hospital, Sichuan University, Chengdu, Sichuan, 610041, ChinaDepartment of Laboratory Medicine, The People's Hospital of Leshan, Leshan, 614000, ChinaDepartment of Laboratory Medicine, West China Hospital, Sichuan University, Chengdu, Sichuan, 610041, ChinaDepartment of Laboratory Medicine, West China Hospital, Sichuan University, Chengdu, Sichuan, 610041, ChinaDepartment of Laboratory Medicine, West China Hospital, Sichuan University, Chengdu, Sichuan, 610041, ChinaDepartment of Laboratory Medicine, West China Hospital, Sichuan University, Chengdu, Sichuan, 610041, China; Corresponding author: Binwu Ying, Department of Laboratory Medicine, West China Hospital, Sichuan University, Chengdu, Sichuan, 610041, ChinaObjectives: Smear-negative pulmonary TB (PTB) is difficult to diagnose. Current diagnosis and treatment monitoring methods have inherent limitations. Droplet digital PCR (ddPCR) is a new technique with high sensitivity. This study presents a novel ddPCR for rapid and sensitive identification of Mycobacterium tuberculosis (MTB). Methods: MTB DNA was detected in respiratory specimens from suspected PTB cases using ddPCR assay, which was directed at two different locations within IS6110. We, for the first time, evaluated the clinical diagnostic ability of this ddPCR for paucibacillary smear-negative PTB. Results: A total of 605 PTB suspects were recruited, including 263 patients with confirmed PTB (84.03% from smear-negative PTB) and 342 without PTB. The sensitivity and specificity of IS6110 ddPCR were 61.22% (95% confidence interval (CI) 55.00-67.10%) and 95.03% (95% CI 92.20-97.10%) for total PTB and 57.92% (95% CI 51.10-64.50%) and 94.57% (95% CI 91.20-96.90%) for smear-negative PTB. ddPCR assay outperformed Xpert MTB/RIF (53.08% vs 28.46%, P = 0.020) in smear-negative PTB detection. Furthermore, effective anti-TB treatment was linked to significantly lower IS6110 copies detected by ddPCR. Conclusion: Herein, we developed and validated a highly sensitive and robust ddPCR assay for MTB quantification in respiratory specimens, which improves diagnosis and therapeutic effect evaluation of smear-negative PTB.http://www.sciencedirect.com/science/article/pii/S1201971222004374TuberculosisPaucibacillarySmear-negative pulmonary tuberculosisDigital droplet polymerase chain reactionXpert MTB/RIF |
| spellingShingle | Zhenzhen Zhao Tao Wu Minjin Wang Xiaojuan Chen Tangyuheng Liu Yanjun Si Yanhong Zhou Binwu Ying A new droplet digital PCR assay: improving detection of paucibacillary smear-negative pulmonary tuberculosis International Journal of Infectious Diseases Tuberculosis Paucibacillary Smear-negative pulmonary tuberculosis Digital droplet polymerase chain reaction Xpert MTB/RIF |
| title | A new droplet digital PCR assay: improving detection of paucibacillary smear-negative pulmonary tuberculosis |
| title_full | A new droplet digital PCR assay: improving detection of paucibacillary smear-negative pulmonary tuberculosis |
| title_fullStr | A new droplet digital PCR assay: improving detection of paucibacillary smear-negative pulmonary tuberculosis |
| title_full_unstemmed | A new droplet digital PCR assay: improving detection of paucibacillary smear-negative pulmonary tuberculosis |
| title_short | A new droplet digital PCR assay: improving detection of paucibacillary smear-negative pulmonary tuberculosis |
| title_sort | new droplet digital pcr assay improving detection of paucibacillary smear negative pulmonary tuberculosis |
| topic | Tuberculosis Paucibacillary Smear-negative pulmonary tuberculosis Digital droplet polymerase chain reaction Xpert MTB/RIF |
| url | http://www.sciencedirect.com/science/article/pii/S1201971222004374 |
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