The performance of an in-house nested-PCR technique for pleural tuberculosis diagnoses

Montenegro, Lílian Maria Lapa; Silva, Bruno César da; Lima, Jaime; Cruz, Heidi Lacerda Alves da; Montenegro, Rosana de Albuquerque; Lundgren, Fernando Luiz Cavalcanti; Filho, Alfredo Pereira Leite de Albuquerque; Schindler, Haiana Charifker

doi:10.1590/0037-8682-0127-2013

Cited by 6 publications

(1 citation statement)

References 27 publications

(46 reference statements)

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“…The sensitivity, specificity, and positive and negative predictive values of PCR in HIV-positive and -negative patients with PTB were 35%, 76.5%, 26.8%, and 73.2%, respectively. In a study on pleural effusion, Montenegro et al (20) observed sensitivities of 84.2%, 72.2%, and 33.3%, respectively, when nested PCR was applied to (i) pleural fluid, blood, and/or urine samples; (ii) blood and/or urine samples without pleural fluid samples; and (iii) pleural fluid samples alone. They concluded that the sensitivity of the test could be improved by the use of different samples.…”

Section: Discussionmentioning

confidence: 99%

Urine-Based Nested PCR for the Diagnosis of Mycobacterium tuberculosis: A Comparative Study Between HIV-Positive and HIV-Negative Patients

Makiani

Davoodian

Baghershiroodi

et al. 2016

Jundishapur J Microbiol

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BackgroundWhile tuberculosis (TB) can be diagnosed by microscopy and culture, the sensitivity of Ziehl-Neelsen staining is variable and culture results require 4 - 8 weeks to be determined. Polymerase chain reaction (PCR) and its modifications, including nested PCR, might be promising methods for the rapid diagnosis of TB.ObjectivesThis study aimed to evaluate the performance of nested PCR on urine samples of human immunodeficiency virus (HIV)-positive and -negative patients with different manifestations of clinical TB.MethodsIn a prospective study, three early-morning urine samples from 100 patients with pulmonary TB (PTB) or extrapulmonary TB (EPTB) were evaluated using a molecular target with insertion element IS6110, specific to the Mycobacterium tuberculosis genome, and nested PCR was performed. The results were analyzed with SPSS version 22.ResultsA total of 100 patients, including 74 (74%) with PTB and 26 (26%) with EPTB, were enrolled. Positive smears were seen in 38 patients (38%). Lymph nodes were the most commonly involved organ in 14 of the 26 (53.8%) EPTB patients (13.5%). Seven (23.1%) of the EPTB patients were HIV-positive. Urine PCR was positive in only 28 patients (28%). Seven HIV-positive patients with PTB showed positive urine PCR results. Moreover, PCR results were positive in only one of the seven HIV-positive subjects with EPTB. Positive PCR results were found in 20 of the 73 HIV-negative patients (27.4%) and in 8 of the 27 HIV-positive patients (29.6%). Therefore, there was no significant difference between the HIV-negative and HIV-positive patients for urine PCR (sensitivity 29.6%, specificity 72.6%; positive and negative predictive values 28% and 72%, respectively; P = 0.138).ConclusionsNested PCR showed the same sensitivity in HIV-positive and HIV-negative patients. It can be applied as a rapid technique for the diagnosis of TB.

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Section: Discussionmentioning

confidence: 99%

Urine-Based Nested PCR for the Diagnosis of Mycobacterium tuberculosis: A Comparative Study Between HIV-Positive and HIV-Negative Patients

Makiani

Davoodian

Baghershiroodi

et al. 2016

Jundishapur J Microbiol

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

Lin

Wang

Lin

et al. 2021

Sci Rep

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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.

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Insight into diagnosis of pleural tuberculosis with special focus on nucleic acid amplification tests

Soni

Guliani

Nehra

et al. 2022

Expert Review of Respiratory Medicine

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The performance of an in-house nested-PCR technique for pleural tuberculosis diagnoses

Cited by 6 publications

References 27 publications

Urine-Based Nested PCR for the Diagnosis of Mycobacterium tuberculosis: A Comparative Study Between HIV-Positive and HIV-Negative Patients

Urine-Based Nested PCR for the Diagnosis of Mycobacterium tuberculosis: A Comparative Study Between HIV-Positive and HIV-Negative Patients

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

Insight into diagnosis of pleural tuberculosis with special focus on nucleic acid amplification tests

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