Multiplex polymerase chain reaction using insertion sequence 6110 (IS6110) and mycobacterial protein fraction from BCG of Rm 0.64 in electrophoresis target genes for diagnosis of tuberculous lymphadenitis

Sharma, Kusum; Gupta, Nalini; Sharma, Aman; Singh, Gagandeep; Gupta, PK; Rajwanshi, Arvind; Varma, Saurabh; Sharma, Megha

doi:10.4103/0255-0857.108714

Cited by 13 publications

(8 citation statements)

References 15 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Various studies using PCR report a variable sensitivity from as low as 30–40% (using single primer) to as high as 80% (multiple primers) for GITB . Studies report no variation in the accuracy of assay irrespective of the presence or absence of granuloma or caseous necrosis in tissue biopsy . Other benefits of PCR include the rapidity in obtaining the results with a quick and accurate diagnosis.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Multiplex Polymerase Chain Reaction for diagnosis of gastrointestinal tuberculosis

et al. 2018

View full text Add to dashboard Cite

Background and Aim To evaluate the role of multiplex polymerase chain reaction (PCR) for diagnosis of gastrointestinal tuberculosis (GITB). Methods This was a prospective observational study conducted from July 2015 to November 2016 at a tertiary care teaching institution in north India. Fifty individuals with clinically suspected GITB and older than 18 years of age were recruited. Patients underwent radiological investigations, esophagogastroduodenoscopy, or colonoscopy as clinically indicated. Multiple biopsies for tissue diagnosis and PCR were taken. All specimens were subjected to Ziehl Neelsen staining, histopathology, and multiplex PCR using specific primers for genes IS6110, MPB64, and Protein b. The performance of the assay was assessed using a composite reference standard for diagnosis of tuberculosis. It comprised a combination of clinical characteristics and microbiological methods, including smear, Bactenecin (BACTEC) culture, histopathology, and response to antitubercular therapy. Results A final diagnosis of tuberculosis was made in 32 cases (Duodenal‐4, Ileo‐colonic‐28). The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of histopathology for the diagnosis of tuberculosis was 28.12, 100, 100, and 43.9%, respectively. The sensitivity, specificity, PPV and NPV of BACTEC Mycobacteria Growth Indicator Tube (MGIT) culture for the diagnosis of tuberculosis was 9.3, 100, 100, and 38.29%, respectively. The sensitivity, specificity, PPV, and NPV of multiplex PCR for the diagnosis of tuberculosis was 87.5, 100, 100, and 86.2%, respectively. Conclusion Multiplex PCR using specific primers for genes IS6110, MPB64, and Protein b had a higher sensitivity compared to conventional techniques for the diagnosis of GITB.

show abstract

Section: Discussionmentioning

confidence: 99%

“…tissue biopsy. [22][23][24] Other benefits of PCR include the rapidity in obtaining the results with a quick and accurate diagnosis. Kulkarni et al used PCR for the identification of M. tuberculosis with a single primer for 340 base pair nucleotide sequence.…”

Section: Discussionmentioning

confidence: 99%

Multiplex Polymerase Chain Reaction for diagnosis of gastrointestinal tuberculosis

et al. 2018

View full text Add to dashboard Cite

show abstract

“…(9.10) Sharma et al found a positivity rate of Zheil Neelson stain,culture and PCR to be 30%,26.3% and 91.3%respectively . (11) Recently,Xpert MTB RIF assay(Cefeid Sunnyvale CA,USA) has been found to have a sensitivity of 81.2% for lymph nodes,62.8% for CSF and 21.4% for pleural fluid . (12) Navarro Viasaro et al observed that urogenital TB was the third most frequent EPTB infection ,the first two being pleural and lymph node TB.…”

Section: Discussionmentioning

confidence: 99%

Truelab Micro PCR In Diagnosis of Extrapulmonary Tuberculosis: Our Experience

Hawaldar¹,

Sodani²

2016

JMRR

View full text Add to dashboard Cite

Introduction: India carries about 25% burden of tuberculosis.It is estimated that about 1520% of all TB cases are of extrapulmonary origin. Recently,molecular amplification methods such as PCR and RT PCR have attracted interest of laboratorians as the diagnosis can be made in a short time and treatment initiated at the proper time.The present study was undertaken to evaluate the role of Truelab MicroPCR (Molbio diagnostics) in diagnosis of EPTB. Materials and Methods: This was a retrospective study conducted between January 2014 to June 2016.Atotal of 447 patients suspected of having EPTB clinically were included in the study. Specimens were collected from patients,both male and female,from different sites like pus,body fluids,lymph node aspirates,CSF,semen,urine ,menstrual blood and endometrial tissue etc.according to standard protocols. All the specimens were run on Truelab MicroPCR system(Molbio diagnostics) according to standard protocols. Results: A total of 447 patients data was used for this study. Maximum number of samples were endometrial tissue (59.9%) followed by lymph node aspirates (11.63%),pus (4.92%),urine (4.47%) , pleural fluid & menstrual blood (3.57%) CSF (3.13%) , Ascitic fluid (2.90%) Synovial fluid (1.1%), vitreous fluid and peritoneal fluid (0.89%) , BAL fluid (0.67%). Out of these different extra pulmonary samples 19(4.25%) were positive by RT PCR and 95.75% were negative. Highest positivity rate was observed inpus(27.2%) , followed by CSF (21.4%) , menstrual blood & pleural fluid (6.25%) ,lymph node aspirates(5.76%), urine(5%)and 1.49% in endometrial tissue. Conclusion: Truelab Micro PCR is portable,easy to use and provides accurate diagnosis of disease like EPTB with a rapid turn around time. .It does not require skilled manpower and elaborate infrastructure and can be easily installed in resource limited settings. A complete patient centric solution for patients suffering from EPTB is a necessity and rapid ,accurate diagnosis with Truelab MicroPCR and prompt therapywill help control TB burden in India.

show abstract

“…Of the samples spot and early morning sputum was collected in 59 patients, 16 were bronchial wash specimen and 5 were tracheal aspirate. A total of 65 EPTB suspected samples like urine, pus, FNAC, pleural fluid, peritoneal fluid, ascitic fluid, Synovial fluid, tissue, pericardial fluid and CSF sent to the Central laboratory were also processed 3 . Consecutive whole early morning urine were collected in sterile container.…”

Section: Materials and Methods Study Design And Populationmentioning

confidence: 99%

“…The chief difficulty with extrapulmonary specimens is the low yield of bacilli and are consequently associated with lower sensitivity in Microscopy and culture. Microscopy is positive in only 10% of EPTB cases whereas culture positivity ranges from 12 to 70% of cases 3 . The positivity rates of Ziehl-Neelsen smear, culture, and multiplex PCR were 30%, 26.3% and 91.3%, respectively, in confirmed tubercular lymphadenitis patients 4 and its diagnosis remains a challenge since granulomatous lymphadenopathy has an extensive differential diagnosis.…”

Section: Introductionmentioning

confidence: 98%

Ergonomic Diagnostic Tool based on Chip Mini RT-PCR for Diagnosis of Pulmonary and Extra Pulmonary Tuberculosis

Mangayarkarasi¹,

Sneka²,

R³

et al. 2019

J Pure Appl Microbiol

View full text Add to dashboard Cite

Diagnosis of Tuberculosis (TB) is a challenging problem with the currently available conventional methods such as microscopy for Acid Fast Bacilli (AFB) and culture techniques.The diagnosis of Extra pulmonary tuberculosis (EPTB) is still more challenging due to the low yield of bacilli in the clinical specimens. Recently, a chip based Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) was introduced in India. This study was conducted for finding out the sensitivity and specificity of TrueNAT RT-PCR for the diagnosis of Pulmonary and EPTB at a Tertiary Care Hospital in Southern India. A total of 145 samples including both Pulmonary (80) and EPTB (65) were examined by Smear microscopy, culture on Lowenstein Jensen (LJ) medium and TruNat RT PCR. All the positive samples were confirmed by conventional PCR technique as well. Out of 80 Pulmonary samples such as sputum, bronchial wash and tracheal aspirate 41(51.25%) samples were positive in RT-PCR, 22 (27.5%) were positive in microscopy and 29 (36.25%) was positive for culture on LJ medium. Among the 65 EPTB samples such as pus, pleural fluid, Cerebro spinal fluid (CSF), ascitic fluid, Tissue, Peritoneal fluid, Pericardial fluid, Urine, synovial fluid, Fine needle aspiration cytology (FNAC) 38 (58.46%) were positive in RT-PCR, 08 (12.30%) were positive in microscopy and 31(47.69%) were positive for culture on Lowenstein Jensen (LJ) medium. The sensitivity and specificity of TrueNat RT-PCR for the diagnosis of Pulmonary tuberculosis (PTB) is 93.1% and 72.5 % and for EPTB is 96.77 %and 76.4 %, respectively. The newer diagnostic tool has a Turn Around Time (TAT) of less than 2 hours, can detect rifampicin resistance, longer shelf life, cost effective and can work from 2°C to 40°C. It is portable and comes with an inbuilt rechargeable battery which makes it a compatible equipment for any health care setup and out reach programs to detect and treat patients even in remote villages.

show abstract

Multiplex polymerase chain reaction using insertion sequence 6110 (IS6110) and mycobacterial protein fraction from BCG of Rm 0.64 in electrophoresis target genes for diagnosis of tuberculous lymphadenitis

Abstract: In conclusion, techniques like MPCR with high sensitivity and specificity can play an important role in rapid diagnosis of TBLA.

Cited by 13 publications

References 15 publications

Multiplex Polymerase Chain Reaction for diagnosis of gastrointestinal tuberculosis

Multiplex Polymerase Chain Reaction for diagnosis of gastrointestinal tuberculosis

Truelab Micro PCR In Diagnosis of Extrapulmonary Tuberculosis: Our Experience

Ergonomic Diagnostic Tool based on Chip Mini RT-PCR for Diagnosis of Pulmonary and Extra Pulmonary Tuberculosis

Contact Info

Product

Resources

About