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Peer-reviewedCC BYJournal of global health

Diagnosis of tuberculous lymphadenopathy: the role of droplet digital PCR.

  • Yu-Feng Wei1,2
  • Ching-Yi Chen1,3
  • Ping-Huai Wang4
  • Yao-Wen Kuo5,6
  • Chin-Chung Shu7,8
  • Bo-Shiun Yan9
9 affiliations
  1. 1School of Medicine for International Students, College of Medicine, I-Shou University, Kaohsiung, Taiwan.
  2. 2Department of Internal Medicine, E-Da Cancer Hospital, Kaohsiung, Taiwan.
  3. 3Division of Chest Medicine, Department of Internal Medicine, E-Da Hospital, Kaohsiung, Taiwan.
  4. 4Department of Internal Medicine, Far Eastern Memorial Hospital, New Taipei City, Taiwan.
  5. 5Department of Integrated Diagnostics & Therapeutics, National Taiwan University Hospital, Taipei, Taiwan.
  6. 6Department of Internal Medicine, College of Medicine, National Taiwan University, Taipei, Taiwan.
  7. 7Department of Internal Medicine, National Taiwan University Hospital, Taipei, Taiwan.
  8. 8Graduate Institute of Clinical Medicine, College of Medicine, National Taiwan University, Taipei, Taiwan.
  9. 9Graduate Institute of Biochemistry and Molecular Biology, College of Medicine, National Taiwan University, Taipei, Taiwan.
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Abstract

Background

Tuberculous lymphadenopathy (TB-LAP) is the most common form of extrapulmonary TB and remains diagnostically challenging due to its paucibacillary nature. We evaluated the diagnostic performance of droplet digital PCR (ddPCR) for detecting Mycobacterium tuberculosis (Mtb) DNA in formalin-fixed paraffin-embedded (FFPE) lymph node specimens.Methods

We retrospectively analysed 102 lymph node FFPE specimens with microbiologically confirmed or pathology-suspected TB-LAP. We performed ddPCR targeting the IS6110 gene and correlated it with pathological findings and mycobacterial culture results. Diagnostic performance was assessed using receiver operating characteristic analysis.Results

Pathological findings included granulomatous inflammation (GI) (n = 67), caseating GI (CGI) (n = 22), CGI with acid-fast bacilli (AFB) (n = 7), GI with AFB (n = 1), and atypical findings (n = 5). There was a graded association between ddPCR and pathological severity (P for trend = 0.002), with the highest levels in CGI plus AFB and the lowest in non-TB-LAP cases. Mtb DNA concentration (copies/μL) was significantly higher in AFB-positive specimens (mean (x̄) = 22.9, standard deviation (SD) = 61.5) compared with AFB-negative specimens (x̄ = 0.6, SD = 0.8) (P = 0.044). Among 52 specimens sent for mycobacterial culture, 9 (17%) were culture-positive, and these specimens had significantly higher ddPCR values (P = 0.025). Receiver operating characteristic analysis yielded an area under the curve of 0.715 (95% confidence interval = 0.536-0.894) for AFB positivity. Using a cutoff value of >1.25 copies/μL resulted in 50% sensitivity and 86% specificity. Lowering the cutoff to >0.33 copies/μL increased sensitivity to 88%, but reduced specificity to 51%.Conclusions

We found that ddPCR quantitatively detects Mtb DNA in lymph node FFPE specimens and correlates with pathological severity and culture positivity. Although the overall diagnostic accuracy was moderate, ddPCR demonstrated distinct sensitivity and specificity profiles at different cutoff values and may serve as a useful adjunctive diagnostic tool when interpreted in conjunction with clinical and histopathological findings in TB-LAP.

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Cite

BibTeXRIS
BibTeX
@article{wei2026diagnosis,
  title = {{Diagnosis of tuberculous lymphadenopathy: the role of droplet digital PCR.}},
  author = {Yu-Feng Wei and Ching-Yi Chen and Ping-Huai Wang and Yao-Wen Kuo and Chin-Chung Shu and Bo-Shiun Yan},
  journal = {Journal of global health},
  year = {2026},
  month = oct,
  doi = {10.7189/jogh.16.04244},
  url = {https://europepmc.org/article/MED/42852602},
  note = {CC BY},
}
RIS
TY  - JOUR
TI  - Diagnosis of tuberculous lymphadenopathy: the role of droplet digital PCR.
AU  - Yu-Feng Wei
AU  - Ching-Yi Chen
AU  - Ping-Huai Wang
AU  - Yao-Wen Kuo
AU  - Chin-Chung Shu
AU  - Bo-Shiun Yan
PY  - 2026
DA  - 2026/10/09
JO  - Journal of global health
DO  - 10.7189/jogh.16.04244
UR  - https://europepmc.org/article/MED/42852602
AB  - Background  Tuberculous lymphadenopathy (TB-LAP) is the most common form of extrapulmonary TB and remains diagnostically challenging due to its paucibacillary nature. We evaluated the diagnostic performance of droplet digital PCR (ddPCR) for detecting Mycobacterium tuberculosis (Mtb) DNA in formalin-fixed paraffin-embedded (FFPE) lymph node specimens.Methods  We retrospectively analysed 102 lymph node FFPE specimens with microbiologically confirmed or pathology-suspected TB-LAP. We performed ddPCR targeting the IS6110 gene and correlated it with pathological findings and mycobacterial culture results. Diagnostic performance was assessed using receiver operating characteristic analysis.Results  Pathological findings included granulomatous inflammation (GI) (n = 67), caseating GI (CGI) (n = 22), CGI with acid-fast bacilli (AFB) (n = 7), GI with AFB (n = 1), and atypical findings (n = 5). There was a graded association between ddPCR and pathological severity (P for trend = 0.002), with the highest levels in CGI plus AFB and the lowest in non-TB-LAP cases. Mtb DNA concentration (copies/μL) was significantly higher in AFB-positive specimens (mean (x̄) = 22.9, standard deviation (SD) = 61.5) compared with AFB-negative specimens (x̄ = 0.6, SD = 0.8) (P = 0.044). Among 52 specimens sent for mycobacterial culture, 9 (17%) were culture-positive, and these specimens had significantly higher ddPCR values (P = 0.025). Receiver operating characteristic analysis yielded an area under the curve of 0.715 (95% confidence interval = 0.536-0.894) for AFB positivity. Using a cutoff value of >1.25 copies/μL resulted in 50% sensitivity and 86% specificity. Lowering the cutoff to >0.33 copies/μL increased sensitivity to 88%, but reduced specificity to 51%.Conclusions  We found that ddPCR quantitatively detects Mtb DNA in lymph node FFPE specimens and correlates with pathological severity and culture positivity. Although the overall diagnostic accuracy was moderate, ddPCR demonstrated distinct sensitivity and specificity profiles at different cutoff values and may serve as a useful adjunctive diagnostic tool when interpreted in conjunction with clinical and histopathological findings in TB-LAP.
SN  - 2047-2978
SN  - 2047-2986
ER  -