Use of conventional PCR and smear microscopy to diagnose pulmonary tuberculosis in the Amazonian rainforest area

Carniel, Francieli; Costa, Elis Regina Dalla; Bello, Graziele Lima; Martins, Cristiana Alves; Scherer, Luciene Cardoso; Rossetti, Maria Lúcia Rosa

doi:10.1590/1414-431x20143899

Cited by 8 publications

(10 citation statements)

References 20 publications

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“…29 In Brazil in 2014, Carniel et al assessed the performance of PCR (IS6110) in DNA extracted from slides prepared for microscopy to diagnose TB at a reference health unit for TB treatment in the Amazonian rainforest area, a difficult to access region with a high TB prevalence. 30 They demonstrated that an in-house PCR using DNA extracted from ZN slides was a feasible alternative method for the detection of M. tuberculosis, with a shorter turnaround time for results and an increased case detection rate. They showed that this method could offer a new approach for accurate TB diagnosis, especially in remote settings where culture is not available.…”

Section: Dna Extracted From Slides For Tb Diagnosismentioning

confidence: 99%

Exploring tuberculosis by molecular tests on DNA isolated from smear microscopy slides

Rakotosamimanana

Rabodoarivelo

Palomino

et al. 2017

International Journal of Infectious Diseases

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Tuberculosis (TB) is an infectious disease of global public health importance caused by Mycobacterium tuberculosis complex. The disease has worsened with the emergence of multidrug-resistant (MDR)-TB strains. The timely diagnosis and treatment of TB remains a key public health priority, and laboratories have a critical role in the rapid and accurate detection of TB and drug resistance. Molecular assays based on nucleic acid amplification techniques have been developed for the rapid, sensitive, and specific diagnosis of TB, with the ability to determine the drug sensitivity status. These molecular techniques are now available or are being implemented in developing countries. However, traditional microscopy and culture methods cannot yet be replaced; the molecular assays can be applied in parallel with these tests for the diagnosis of TB or for drug susceptibility testing. Performing such molecular tests is often restricted by constraints with regard to sputum sample storage and safe transportation from remote health centres to central laboratories. Since smear slides are performed routinely for the diagnosis of TB in most TB diagnostic laboratories, they are readily available and could be the ideal tool to transport sputum for further molecular tests. The aim of this review was to provide a comprehensive survey on the use of smear slides for both TB diagnosis and the molecular test approach. Based on the literature, stained smear microscopy slides can be a safe system for the transportation of sputum specimens from remote health centres to reference TB laboratories for further molecular TB or MDR-TB detection, and could help in the rapid diagnosis and therefore timely management of TB patients.

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Section: Dna Extracted From Slides For Tb Diagnosismentioning

confidence: 99%

Exploring tuberculosis by molecular tests on DNA isolated from smear microscopy slides

Rakotosamimanana

Rabodoarivelo

Palomino

et al. 2017

International Journal of Infectious Diseases

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“…We found the presence of M. tuberculosis DNA in patient sample 1 (Table II), however, the presence of the bacillus was not visualised on microscopy. (28) Combining the FN-stained slide with bacillus DNA detection and drug resistance testing increases the effectiveness of the analyses and makes them more cost-effective. (27) Evaluating the results, genotyping analyses in M. tuberculosis strains could contribute significantly to disease control, indicate possible epidemiological associations, detect suspicious outbreak information and cross-contamination in laboratories, and even distinguish between reinfections exogenous and endogenous reactivation during retreatment.…”

Section: Discussionmentioning

confidence: 99%

Rapid detection of Mycobacterium tuberculosis DNA and genetic markers for Isoniazid resistance in Ziehl-Neelsen stained slides

Bello

Morais

Jesus

et al. 2020

Mem. Inst. Oswaldo Cruz

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BACKGROUND Early diagnosis of tuberculosis (TB) and identification of strains of Mycobacterium tuberculosis resistant to anti-TB drugs are considered the main factors for disease control. OBJECTIVES To standardise a real-time polymerase chain reaction (qPCR) assay technique and apply it to identify mutations involved in M. tuberculosis resistance to Isoniazid (INH) directly in Ziehl-Neelsen (ZN) stained slides. METHODS Were analysed 55 independent DNA samples extracted from clinical isolates of M. tuberculosis by sequencing. For application in TB diagnosis resistance, 59 ZN-stained slides were used. The sensitivity, specificity and Kappa index, with a 95% confidence interval (CI95%), were determined. FINDINGS The agreement between the tests was, for the katG target, the Kappa index of 0.89 (CI95%: 0.7-1.0). The sensitivity and specificity were 97.6% (CI95%: 87.7-99.9) and 91.7% (CI95%: 61.5-99.5), respectively. For inhA, the Kappa index was 0.92 (CI95%: 0.8-1.0), the sensitivity and specificity were 94.4% (CI95%: 72.7-99.8) and 97.3% (CI95%: 85.8-99.9), respectively. The use of ZNstained slides for drug-resistant TB detection showed significant results when compared to other standard tests for drug resistance. MAIN CONCLUSIONS qPCR genotyping proved to be an efficient method to detect genes that confer M. tuberculosis resistance to INH. Thus, qPCR genotyping may be an alternative instead of sequencing.

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“…Nos résultats corroborent ceux d’autres auteurs notamment Geleta et al en Ethiopie [ 11 ], Giang et al au Vietnam [ 12 ] et Lawn et al en Afrique du Sud [ 13 ]. Pour certains auteurs, Le couple ZN / PCR conventionnelle permet de détecter plus de cas de TBC que le ZN seul [ 14 ]. Notre étude a aussi montré une sensibilité élevée de GeneXpert MTB/RIF dans la détection de la TBC chez les personnes vivant avec le VIH en particulier corroborant ainsi d’autres études [ 1 , 15 ].…”

Section: Discussionunclassified

GeneXpert MTB/RIF dans le dépistage de la tuberculose pulmonaire à l’Hôpital Provincial Général de Référence de Bukavu, à l’Est de la République Démocratique du Congo: quelles leçons tirées après 10 mois d’utilisation?

Lupande¹,

Kaishusha²,

Mihigo³

et al. 2017

Pan Afr Med J

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IntroductionEn Afrique subsaharienne, les méthodes de diagnostic de la tuberculose sont insuffisantes et reposent essentiellement sur la microscopie. Elles constituent un réel frein pour le contrôle de la tuberculose. La présente étude voudrait évaluer les performances du GeneXpert MTB/RIF vis à vis de la microscopie classique de Ziehl-Neelsen à l’Hôpital Provincial Général de Référence de Bukavu, à l’Est de la République Démocratique du Congo après 10 mois d’utilisation.MéthodesLes résultats de la coloration au Ziehl-Neelsen et de la biologie moléculaire sur GeneXpert MTB/RIF de 452 patients suspects de tuberculose ont été colligés. La validité d’un test par rapport à l’autre dans la détection de la tuberculose a été étudiée.RésultatsDans le groupe entier, la fréquence de la tuberculose pulmonaire était de 16.3%. La positivité était significativement plus élevée pour le GeneXpert MTB/RIF que pour le Ziehl-Neelsen dans le groupe entier (15.9% vs 9.3%, p= 0.03) et chez les séropositifs pour le VIH (52.0% vs 24.0%; p = 0.007). Cependant, la sensibilité de GeneXpert MTB/RIF comparé au Ziehl-Neelsen n’était pas maximale (95.2%). Enfin, GeneXpert MTB/RIF a détecté 20.8% de résistance à la rifampicine.ConclusionLa présente étude confirme la supériorité de GeneXpert MTB/RIF sur la coloration de Ziehl-Neelsen dans la détection de la tuberculose et dans la prédiction de la multi résistance. Son utilisation systématique couplée au Ziehl-Neelsen permettrait de mieux contrôler la tuberculose en Afrique subSaharienne.

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Use of conventional PCR and smear microscopy to diagnose pulmonary tuberculosis in the Amazonian rainforest area

Cited by 8 publications

References 20 publications

Exploring tuberculosis by molecular tests on DNA isolated from smear microscopy slides

Exploring tuberculosis by molecular tests on DNA isolated from smear microscopy slides

Rapid detection of Mycobacterium tuberculosis DNA and genetic markers for Isoniazid resistance in Ziehl-Neelsen stained slides

GeneXpert MTB/RIF dans le dépistage de la tuberculose pulmonaire à l’Hôpital Provincial Général de Référence de Bukavu, à l’Est de la République Démocratique du Congo: quelles leçons tirées après 10 mois d’utilisation?

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