Development of a two-step nucleic acid amplification test for accurate diagnosis of the Mycobacterium tuberculosis complex

Abstract The Mycobacterium tuberculosis complex (MTBC) remains one of the top 10 leading causes of death globally. The early diagnosis of MTBC can reduce mortality and mitigate disease transmission. However, current nucleic acid amplification diagnostic test methods are generally time-consuming and...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Chien-Ru Lin, Hsin-Yao Wang, Ting-Wei Lin, Jang-Jih Lu, Jason Chia-Hsun Hsieh, Min-Hsien Wu
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/038efb30330144649b6896e7670b9cd7
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:038efb30330144649b6896e7670b9cd7
record_format	dspace
spelling	oai:doaj.org-article:038efb30330144649b6896e7670b9cd72021-12-02T13:34:51ZDevelopment of a two-step nucleic acid amplification test for accurate diagnosis of the Mycobacterium tuberculosis complex10.1038/s41598-021-85160-22045-2322https://doaj.org/article/038efb30330144649b6896e7670b9cd72021-03-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-85160-2https://doaj.org/toc/2045-2322Abstract The Mycobacterium tuberculosis complex (MTBC) remains one of the top 10 leading causes of death globally. The early diagnosis of MTBC can reduce mortality and mitigate disease transmission. However, current nucleic acid amplification diagnostic test methods are generally time-consuming and show suboptimal diagnostic performance, especially in extrapulmonary MTBC samples or acid-fast stain (AFS)-negative cases. Thus, development of an accurate assay for the diagnosis of MTBC is necessary, particularly under the above mentioned conditions. In this study, a single-tube nested real-time PCR assay (N-RTP) was developed and compared with a newly in-house-developed high-sensitivity real-time PCR assay (HS-RTP) using 134 clinical specimens (including 73 pulmonary and 61 extrapulmonary specimens). The amplification efficiency of HS-RTP and N-RTP was 99.8% and 100.7%, respectively. The sensitivity and specificity of HS-RTP and N-RTP for the diagnosis of MTBC in these specimens were 97.5% (77/79) versus 94.9% (75/79) and 80.0% (44/55) versus 89.1% (49/55), respectively. The sensitivity and specificity of HS-RTP and N-RTP for the diagnosis of MTBC in pulmonary specimens were 96.3% (52/54) versus 96.3% (52/54) and 73.7.0% (14/19) versus 89.5% (17/19), respectively; in extrapulmonary specimens, the sensitivity and specificity of HS-RTP and N-RTP were 100% (25/25) versus 92% (23/25) and 83.3% (30/36) versus 88.9% (32/36), respectively. Among the AFS-negative cases, the sensitivity and specificity of HS-RTP and N-RTP were 97.0% (32/33) versus 90.9% (30/33) and 88.0% (44/50) versus 92.0% (46/50), respectively. Overall, the sensitivity of HS-RTP was higher than that of N-RTP, and the performance was not compromised in extrapulmonary specimens and under AFS-negative conditions. In contrast, the specificity of the N-RTP assay was higher than that of the HS-RTP assay in all types of specimens. In conclusion, the HS-RTP assay would be useful for screening patients suspected of exhibiting an MTBC infection due to its higher sensitivity, while the N-RTP assay could be used for confirmation because of its higher specificity. Our results provide a two-step method (screen to confirm) that simultaneously achieves high sensitivity and specificity in the diagnosis of MTBC.Chien-Ru LinHsin-Yao WangTing-Wei LinJang-Jih LuJason Chia-Hsun HsiehMin-Hsien WuNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-11 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Chien-Ru Lin Hsin-Yao Wang Ting-Wei Lin Jang-Jih Lu Jason Chia-Hsun Hsieh Min-Hsien Wu Development of a two-step nucleic acid amplification test for accurate diagnosis of the Mycobacterium tuberculosis complex
description	Abstract The Mycobacterium tuberculosis complex (MTBC) remains one of the top 10 leading causes of death globally. The early diagnosis of MTBC can reduce mortality and mitigate disease transmission. However, current nucleic acid amplification diagnostic test methods are generally time-consuming and show suboptimal diagnostic performance, especially in extrapulmonary MTBC samples or acid-fast stain (AFS)-negative cases. Thus, development of an accurate assay for the diagnosis of MTBC is necessary, particularly under the above mentioned conditions. In this study, a single-tube nested real-time PCR assay (N-RTP) was developed and compared with a newly in-house-developed high-sensitivity real-time PCR assay (HS-RTP) using 134 clinical specimens (including 73 pulmonary and 61 extrapulmonary specimens). The amplification efficiency of HS-RTP and N-RTP was 99.8% and 100.7%, respectively. The sensitivity and specificity of HS-RTP and N-RTP for the diagnosis of MTBC in these specimens were 97.5% (77/79) versus 94.9% (75/79) and 80.0% (44/55) versus 89.1% (49/55), respectively. The sensitivity and specificity of HS-RTP and N-RTP for the diagnosis of MTBC in pulmonary specimens were 96.3% (52/54) versus 96.3% (52/54) and 73.7.0% (14/19) versus 89.5% (17/19), respectively; in extrapulmonary specimens, the sensitivity and specificity of HS-RTP and N-RTP were 100% (25/25) versus 92% (23/25) and 83.3% (30/36) versus 88.9% (32/36), respectively. Among the AFS-negative cases, the sensitivity and specificity of HS-RTP and N-RTP were 97.0% (32/33) versus 90.9% (30/33) and 88.0% (44/50) versus 92.0% (46/50), respectively. Overall, the sensitivity of HS-RTP was higher than that of N-RTP, and the performance was not compromised in extrapulmonary specimens and under AFS-negative conditions. In contrast, the specificity of the N-RTP assay was higher than that of the HS-RTP assay in all types of specimens. In conclusion, the HS-RTP assay would be useful for screening patients suspected of exhibiting an MTBC infection due to its higher sensitivity, while the N-RTP assay could be used for confirmation because of its higher specificity. Our results provide a two-step method (screen to confirm) that simultaneously achieves high sensitivity and specificity in the diagnosis of MTBC.
format	article
author	Chien-Ru Lin Hsin-Yao Wang Ting-Wei Lin Jang-Jih Lu Jason Chia-Hsun Hsieh Min-Hsien Wu
author_facet	Chien-Ru Lin Hsin-Yao Wang Ting-Wei Lin Jang-Jih Lu Jason Chia-Hsun Hsieh Min-Hsien Wu
author_sort	Chien-Ru Lin
title	Development of a two-step nucleic acid amplification test for accurate diagnosis of the Mycobacterium tuberculosis complex
title_short	Development of a two-step nucleic acid amplification test for accurate diagnosis of the Mycobacterium tuberculosis complex
title_full	Development of a two-step nucleic acid amplification test for accurate diagnosis of the Mycobacterium tuberculosis complex
title_fullStr	Development of a two-step nucleic acid amplification test for accurate diagnosis of the Mycobacterium tuberculosis complex
title_full_unstemmed	Development of a two-step nucleic acid amplification test for accurate diagnosis of the Mycobacterium tuberculosis complex
title_sort	development of a two-step nucleic acid amplification test for accurate diagnosis of the mycobacterium tuberculosis complex
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/038efb30330144649b6896e7670b9cd7
work_keys_str_mv	AT chienrulin developmentofatwostepnucleicacidamplificationtestforaccuratediagnosisofthemycobacteriumtuberculosiscomplex AT hsinyaowang developmentofatwostepnucleicacidamplificationtestforaccuratediagnosisofthemycobacteriumtuberculosiscomplex AT tingweilin developmentofatwostepnucleicacidamplificationtestforaccuratediagnosisofthemycobacteriumtuberculosiscomplex AT jangjihlu developmentofatwostepnucleicacidamplificationtestforaccuratediagnosisofthemycobacteriumtuberculosiscomplex AT jasonchiahsunhsieh developmentofatwostepnucleicacidamplificationtestforaccuratediagnosisofthemycobacteriumtuberculosiscomplex AT minhsienwu developmentofatwostepnucleicacidamplificationtestforaccuratediagnosisofthemycobacteriumtuberculosiscomplex
_version_	1718392714541662208

Development of a two-step nucleic acid amplification test for accurate diagnosis of the Mycobacterium tuberculosis complex

Ejemplares similares