Evaluation of multiplex PCR using MPB64 and IS6110 primers for rapid diagnosis of tuberculous meningitis

Lekhak, Sunil; Sharma, Laxmi; Rajbhandari, Reema; Rajbhandari, Pravesh; Shrestha, Resha; Pant, Basant

doi:10.1016/j.tube.2016.05.016

Cited by 23 publications

(13 citation statements)

References 27 publications

(27 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…PCR for MTB has also been used in diagnosis of extrapulmonary tuberculosis including abdominal tuberculosis and tuberculous meningitis. [10][11][12] PCR for MTB has been very useful in differentiating Mycobacterium tuberculosis )MTB( infection from Nontuberculous mycobacteria due to use of DNA sequence IS6110 specific to MTB. PCR based NAAT has also been found to be a better diagnostic tool than microscopy and culture in pediatric pulmonary tuberculosis.…”

Section: Resultsmentioning

confidence: 99%

Comparison of real time PCR with phenotypic methods in bronchoalveolar lavage in diagnosis of sputum smear negative pulmonary tuberculosis patients

et al. 2018

View full text Add to dashboard Cite

Background: Early diagnosis of pulmonary tuberculosis is of utmost importance for proper control of the disease in the patient. Diagnosis of pulmonary tuberculosis is usually by acid fast bacilli (AFB) smear examination and culture Mycobacterium tuberculosis (MTB). In this study, we have employed polymerase chain reaction (PCR) for MTB in bronchoalveolar lavage (BAL) along with AFB smear and culture MTB for early diagnosis of pulmonary tuberculosis.Methods: A prospective observational study was conducted in the Department of Pulmonary Medicine, Era’s Lucknow medical college and Hospital, Lucknow over a period of two years. A total of 123 previously treated cases of pulmonary tuberculosis were enrolled for the study whose two sputum smear samples were negative for AFB. These patients underwent fibreoptic bronchoscopy and BAL was obtained which was sent for AFB smear, culture MTB and PCR for MTB.Results: The examination of BAL revealed the highest sensitivity for culture MTB at 87.4% followed by PCR for MTB at 73.8% and then AFB smear at 61.2%. PCR for MTB helped in diagnosing an additional 12% patients of pulmonary tuberculosis which were negative on AFB smear and an additional 6.8% patients which were negative on culture MTB.Conclusions: PCR for MTB is useful in making an early diagnosis of pulmonary tuberculosis especially in paucibacillary cases negative on AFB smear and also in some culture MTB negative patients.

show abstract

Section: Resultsmentioning

confidence: 99%

Comparison of real time PCR with phenotypic methods in bronchoalveolar lavage in diagnosis of sputum smear negative pulmonary tuberculosis patients

et al. 2018

View full text Add to dashboard Cite

show abstract

“…Therefore, in this research, we added another target gene, the mpb64 gene. The mpb64 is encoded in the RD2 region of the M. tuberculosis genome [25], which creates the MPB64 protein, with a strong speci city for MTBC [25,26]. While, there is some evidence for absence of the mpb64 gene in some substrains of Mycobacterium bovis BCG [27].…”

Section: Discussionmentioning

confidence: 99%

Two Target Genes Based Multiple Cross Displacement Amplification Combined with Lateral Flow Biosensor for the Detection of Mycobacrterium Tuberculosis Complex

Huang

Xiao

Yang

et al. 2020

Preprint

View full text Add to dashboard Cite

Background: Mycobacterium tuberculosis complex (MTBC) causes tuberculosis (TB), which is a global public health problem that seriously endangers public health. Hence, development of a new and rapid method to detect MTBC is of great significance for prevention and treatment of TB. Results: In this study, a multiple cross displacement amplification combined with nanoparticle-based lateral flow biosensor (MCDA-LFB) was developed to simultaneously detect two target genes (IS6110 and mpb64) of MTBC. One suit of specific multiple cross displacement amplification (MCDA) primers, which was designed for IS6110 and mpb64 gene, respectively, was validated through using the genomic DNA extracted from reference strain H37Rv. The MCDA products were analyzed using real-time turbidity curve, colorimetric indicator (Malachite Green, MG) and LFB. The conditions of amplification temperature and time were optimized and the established MCDA-LFB method was applied to detect the sputum specimens and MTBC strains from clinical samples. The results show that two sets of MCDA primers for the IS6110 and mpb64 genes have detected MTBC validly. The MCDA reaction conditions were optimized at 67 °C for 35 min. The limit of detection of MCDA assay based on IS6110 and mpb64 genes was 100 fg of genomic DNA template per reaction. The specificity of MCDA-LFB detection was 100%, and no cross-reactions for non-MTBC strains detection. The positive rate of MCDA-LFB for the detection of MTBC strains was equal to that of semi-nested automatic real-time PCR (Xpert MTB/RIF), and had a higher positive rate than acid-fast staining (AFS) when used for the detection of sputum samples. The whole procedure of MCDA-LFB, including genomic template preparation, MCDA reaction and products analysis, was completed with 70 min.Conclusion: The simplicity, rapidity, sensitivity and reliability of the MCDA-LFB based on IS6110 and mpb64 gene of MTBC developed in this study make it potentially significant for the prevention and treatment of TB.

show abstract

“…CB-NAAT has also been shown to be useful in diagnosing abdominal tuberculosis and meningeal tuberculosis. 12,13 A cut off value of >40U/L is usually used to strongly suspect tubercular etiology in a pleural effusion. 14,15 The sensitivity of ADA for diagnosing tubercular pleural effusion has been high and has been reported to be around 90%.…”

Section: Discussionmentioning

confidence: 99%

The advantage of PCR for MTB in comparison to ADA in diagnosing tubercular pleural effusion

et al. 2018

View full text Add to dashboard Cite

Background: Tuberculosis continues to be an important health problem globally. The bacteriological confirmation of diagnosis in extrapulmonary tuberculosis patients is more difficult because most of the cases of extrapulmonary tuberculosis are paucibacillary in nature. In this study we have compared the pleural fluid ADA levels with PCR for MTB in pleural fluid to confirm the diagnosis of tuberculosis in the pleural fluid.Methods: The study was done over two years and a total of 106 patients with a clinico-radiological diagnosis of pleural effusion were enrolled for the study. The pleural fluid was aspirated and examined for total cell count, differential cell count, protein, sugar, ADA and PCR for MTB.A CT Thorax was done in all the 106 patients of pleural effusion and underlying consolidation along with pleural effusion was found in 60 patients.Results: The pleural fluid was exudative in nature in all the patients. 90 patients (84.9%) had lymphocyte predominant pleural effusion while 16 patients (15.1%) had neutrophil predominant pleural effusion. The overall sensitivity of ADA in all the cases of pleural effusion was 85.2% while the overall sensitivity of PCR for MTB in all the cases of pleural effusion was 51.1%. However, in the 60 patients of pleural effusion with underlying lung consolidation, the overall sensitivity of ADA was 69.1% while the overall sensitivity of PCR for MTB was 92.8% for diagnosing tubercular pleural effusion.Conclusions: PCR for MTB is a useful test along with ADA for diagnosing tubercular pleural effusion. PCR for MTB is especially useful in the diagnosis of tubercular pleural effusion in patients with underlying lung consolidation.

show abstract

Evaluation of multiplex PCR using MPB64 and IS6110 primers for rapid diagnosis of tuberculous meningitis

Cited by 23 publications

References 27 publications

Comparison of real time PCR with phenotypic methods in bronchoalveolar lavage in diagnosis of sputum smear negative pulmonary tuberculosis patients

Comparison of real time PCR with phenotypic methods in bronchoalveolar lavage in diagnosis of sputum smear negative pulmonary tuberculosis patients

Two Target Genes Based Multiple Cross Displacement Amplification Combined with Lateral Flow Biosensor for the Detection of Mycobacrterium Tuberculosis Complex

The advantage of PCR for MTB in comparison to ADA in diagnosing tubercular pleural effusion

Contact Info

Product

Resources

About