Rapid real-time PCR for detection of<i>Mycobacterium tuberculosis</i>complex DNA in formalin-fixed paraffin embedded tissues: 16% of histological ‘sarcoid’ may contain such DNA

Surat, Güzin; Wallace, William; Laurenson, Ian F.; Seagar, Amie-Louise

doi:10.1136/jclinpath-2014-202307

Cited by 15 publications

(7 citation statements)

References 29 publications

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“…Nucleic acid-based techniques are rapid and important molecular tools for the pathological diagnosis of TB and include in situ hybridization, qPCR amplification probe methods, PCR-based Xpert MTB/RIF methods, and sequencing (Nyendak et al, 2009). TB IS6110 gene real-time PCR using extracted DNA has been reported to be a sensitive, specific, and rapid method for TB detection in FFPE specimens (Surat et al, 2014). Seo et al evaluated the efficacy of different PCR methods for detecting MTB in FFPE tissues and reported that real-time PCR and nested PCR (N-PCR) can effectively identify MTB in FFPE material with sensitivities of 80.0% and 87.5%, respectively (Seo et al, 2014).…”

Section: Discussionmentioning

confidence: 99%

Using droplet digital PCR in the detection of Mycobacterium tuberculosis DNA in FFPE samples

Cao¹,

Wu²,

Wei

et al. 2020

International Journal of Infectious Diseases

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Background: Droplet digital PCR (ddPCR) is a technology that has higher sensitivity than real-time PCR for the identification of trace DNA. However, the use of ddPCR for the detection of Mycobacterium tuberculosis DNA in pathological samples has not been fully studied. Methods: A total of 65 formalin-fixed and paraffin-embedded (FFPE) specimens were included in this study. Twenty samples with definite results for tuberculosis (TB) were used to establish the ddPCR system for TB detection. ddPCR was then conducted to detect TB DNA in the 45 patients who were classified as 'possible TB' (real-time PCR results in the gray area, Ziehl-Neelsen staining-negative, and hematoxylin and eosin staining showing morphology suspicious for TB). The clinical treatment and disease outcomes were followed to assess the accuracy of ddPCR in the detection of TB DNA. Results: Among the 45 possible TB samples, 26 were ddPCR-positive, 12 were ddPCR-negative, and seven were in the gray area. ddPCR improved the positive rate of 57.8% (26/45) for the samples that were in the gray area by real-time PCR. Moreover, several patients received anti-TB therapy, and the effective ratio of therapy for the ddPCR-positive, ddPCR-negative, and ddPCR-gray area cases was 61.9% (13/21), 50.0% (2/4), and 33.3% (1/3), respectively. Conclusions: ddPCR is more sensitive for detecting mild TB via FFPE samples than real-time PCR. The ddPCR method is of additional value in the diagnosis of TB from pathological samples.

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Section: Discussionmentioning

confidence: 99%

Using droplet digital PCR in the detection of Mycobacterium tuberculosis DNA in FFPE samples

Cao¹,

Wu²,

Wei

et al. 2020

International Journal of Infectious Diseases

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“…However, the broad differential diagnosis of granulomatous diseases and the low sensitivity of ZN staining are major limitations 6 7. Molecular tests detecting Mycobacterium tuberculosis -specific DNA fragment in formalin-fixed paraffin-embedded (FFPE) tissues are reported to have high sensitivity and specificity 8 9. However, most pathological laboratories in developing countries have difficulties in carrying out molecular tests owing to a lack of equipment, qualified PCR laboratories and trained technicians.…”

Section: Introductionmentioning

confidence: 99%

Double staining of bacilli and antigen Ag85B improves the accuracy of the pathological diagnosis of pulmonary tuberculosis

Che

Zhang

et al. 2015

J Clin Pathol

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“…The nucleic acid-based technique provided rapid and important molecular tools in pathological measurement for TB diagnosis, such as in situ hybridization, qPCR amplification probe methods, PCR-based Xpert MTB/RIF methods, and sequencing [16] . TB IS6110 gene real-time PCR using extracted DNA was reported to be a sensitive, specific and rapid method for TB detection in FFPE specimens [17] . Seo AN evaluated the efficacy of different PCR methods for detecting TB in FFPE tissues and implied RT-PCR and N-PCR can effectively identify TB in FFPE material with the sensitivity 80.0% and 87.5%, respectively [18] .…”

Section: Discussionmentioning

confidence: 99%

Using of Digital Droplet PCR in the detection of Mycobacterium tuberculosis DNA with FFPE and Cytological samples

Cao

Wei

et al. 2020

Preprint

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Backgrounds: Accurate diagnosis for TB is essential for TB control. Droplet digital PCR (ddPCR) is a technology that has high sensitivity. However, the use of ddPCR for the detection of TB pathogen in pathological samples is not been fully studied.Methods: A total of 88 samples from the patients who were suspected of tuberculosis were involved in this study, including 65 formalin-fixed and paraffin-embedded (FFPE) specimens and 22 cytological samples. Digital Droplet PCR was used to compare the sensitivity with Real-time PCR. The real-time PCR ct value and ddPCR TB abundant ratio were analyzed by SPSS software.Results: Among the 62 samples that were “possible TB” detected by real-time PCR, 34 samples were ddPCR-positive, 18 samples were ddPCR-negative, and 10 samples were in “gray area” by ddPCR. 26 patients that were ddPCR-positive received anti-tuberculosis therapy and 14 cases of them benefit from the treatment.Conclusions: ddPCR is more sensitive in the detection of TB than Real-time PCR. ddPCR methods can be used as an additional means for the diagnosis of TB from pathological samples.

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Rapid real-time PCR for detection ofMycobacterium tuberculosiscomplex DNA in formalin-fixed paraffin embedded tissues: 16% of histological ‘sarcoid’ may contain such DNA

Cited by 15 publications

References 29 publications

Using droplet digital PCR in the detection of Mycobacterium tuberculosis DNA in FFPE samples

Using droplet digital PCR in the detection of Mycobacterium tuberculosis DNA in FFPE samples

Double staining of bacilli and antigen Ag85B improves the accuracy of the pathological diagnosis of pulmonary tuberculosis

Using of Digital Droplet PCR in the detection of Mycobacterium tuberculosis DNA with FFPE and Cytological samples

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