Nucleic Acid Amplification Testing and Sequencing Combined with Acid-Fast Staining in Needle Biopsy Lung Tissues for the Diagnosis of Smear-Negative Pulmonary Tuberculosis
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BACKGROUND: Smear-negative pulmonary tuberculosis (PTB) is common and difficult to diagnose. In this study, we investigated the diagnostic value of nucleic acid amplification testing and sequencing combined with acid-fast bacteria (AFB) staining of needle biopsy lung tissues for patients with suspected smear-negative PTB. METHODS: Patients with suspected smear-negative PTB who underwent percutaneous transthoracic needle biopsy between May 1, 2012, and June 30, 2015, were enrolled in this retrospective study. Patients with AFB in sputum smears were excluded. All lung biopsy specimens were fixed in formalin, embedded in paraffin, and subjected to acid-fast staining and tuberculous polymerase chain reaction (TB-PCR). For patients with positive AFB and negative TB-PCR results in lung tissues, probe assays and 16S rRNA sequencing were used for identification of nontuberculous mycobacteria (NTM). The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and diagnostic accuracy of PCR and AFB staining were calculated separately and in combination. RESULTS: Among the 220 eligible patients, 133 were diagnosed with TB (men/women: 76/57; age range: 17-80 years, confirmed TB: 9, probable TB: 124). Forty-eight patients who were diagnosed with other specific diseases were assigned as negative controls, and 39 patients with indeterminate final diagnosis were excluded from statistical analysis. The sensitivity, specificity, PPV, NPV, and accuracy of histological AFB (HAFB) for the diagnosis of smear-negative were 61.7% (82/133), 100% (48/48), 100% (82/82), 48.5% (48/181), and 71.8% (130/181), respectively. The sensitivity, specificity, PPV, and NPV of histological PCR were 89.5% (119/133), 95.8% (46/48), 98.3% (119/121), and 76.7% (46/60), respectively, demonstrating that histological PCR had significantly higher accuracy (91.2% [165/181]) than histological acid-fast staining (71.8% [130/181]), P < 0.001. Parallel testing of histological AFB staining and PCR showed the sensitivity, specificity, PPV, NPV, and accuracy to be 94.0% (125/133), 95.8% (46/48), 98.4% (125/127), 85.2% (46/54), and 94.5% (171/181), respectively. Among patients with positive AFB and negative PCR results in lung tissue specimens, two were diagnosed with NTM infections (Mycobacterium avium-intracellulare complex and Mycobacterium kansasii). CONCLUSION: Nucleic acid amplification testing combined with acid-fast staining in lung biopsy tissues can lead to early and accurate diagnosis in patients with smear-negative pulmonary tuberculosis. For patients with positive histological AFB and negative tuberculous PCR results in lung tissue, NTM infection should be suspected and could be identified by specific probe assays or 16S rRNA sequencing.
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Combination of data types, especially the sum of low diagnostics data, provides a multi-level overview, better understanding of complex multifactorial conditions, and leads to faster diagnostics [78,79].
“The combination of data types, especially the sum of low diagnostics data, provides a multi-level overview and better understanding of complex multifactorial conditions, and also leads to a faster diagnostics [78, 79].”
From Converging blockchain and next-generation artificial intelligence technologies to decentralize and accelerate biomedical research and healthcare (2018) · cites this paper as [78]
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- Authors
- Jiang, F., Huang, W., Wang, Y., Tian, P., Chen, X., & Liang, Z.
- Affiliations
- Sichuan University (CN)
- Corresponding author
- Liang, Zongan
- Venue
- PLoS ONE
- Publisher
- Public Library of Science (PLoS)
- Published
- 2016-12-02
- Volume
- 11
- Issue
- 12
- Pages
- e0167342
- ISSN
- 1932-6203 (ISSN-L)
- PMC
- PMC5135092
- OpenAlex
- W2557757021
- Cited by
- 14
- References
- 31
- Citations by year
- 2025: 1 · 2024: 1 · 2023: 1 · 2022: 2 · 2021: 1 · 2020: 1 · 2019: 2 · 2018: 4 · 2017: 1
- FWCI
- 2.15
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- PLoS ONE (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Figshare (submittedVersion, cc-by)
- Topics
- Health Sciences › Medicine › Infectious Diseases › Tuberculosis Research and Epidemiology
- Health Sciences › Medicine › Epidemiology › Mycobacterium research and diagnosis
- Health Sciences › Medicine › Surgery › Infectious Diseases and Tuberculosis
- Keywords
- smear-negative tuberculosis, acid-fast staining, nontuberculous mycobacteria, 16S rRNA gene sequencing, transthoracic needle biopsy, diagnostic accuracy, nontuberculous mycobacterial infection, Mycobacterium avium complex, Mycobacterium kansasii
- MeSH terms
- Adolescent, Adult, Aged, Aged, 80 and over, Female, Humans, Lung, Male, Middle Aged, Mycobacterium tuberculosis, RNA, Bacterial, RNA, Ribosomal, 16S, Staining and Labeling, Tuberculosis, Pulmonary, Polymerase Chain Reaction, Mycobacterium kansasii, Biopsy, Fine-Needle
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