scholarly journals Effect of specimen processing, growth supplement, and different metabolic population on Mycobacterium tuberculosis laboratory diagnosis

PLoS ONE ◽  
2020 ◽  
Vol 15 (4) ◽  
pp. e0230927
Author(s):  
Shashikant Srivastava ◽  
Moti Chapagain ◽  
Tawanda Gumbo
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Laboratory Diagnosis ◽  
Growth Supplement ◽  
Specimen Processing

scholarly journals Tuberculous Meningitis: The Microbiological Laboratory Diagnosis and Its Drug Sensitivity Patterns

2019 ◽  
Vol 3 (2) ◽  
pp. 35
Author(s):  
Titiek Sulistyowati ◽  
Deby Kusumaningrum ◽  
Eko Budi Koendhori ◽  
Ni Made Mertaniasih
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Tuberculous Meningitis ◽  
Drug Sensitivity ◽  
Mycobacterium Tuberculosis Complex ◽  
Laboratory Diagnosis ◽  
Clinical Microbiology Laboratory ◽  
Tuberculosis Complex ◽  
First Line ◽  
Sensitivity Pattern ◽  
Microbiological Laboratory

Background: Tuberculosis continues one of the major challenges to global health. Mycobacterium tuberculosis complex can affect any organ other than the lung parenchyma, include central nervous system. The mortality rate of tuberculous meningitis (TBM) are high worldwide with up to half of survivors suffering irreversible sequelae. Diagnosis of TBM is difficult due to paucibacillary, various clinical manifestation, and invasive procedure to appropriate specimens. Objective: The objectiveis to study the positivity rate of microbiological laboratory diagnosis and its drug sensitivity patterns of TBM patients in Dr. Soetomo Hospital Surabaya during October 2015 until September 2016. Methods: Specimens were cerebrospinal fluids. Identification and drug anti TB sensitivity test were done by BACTEC MGIT 960 system in Clinical Microbiology Laboratory Dr. Soetomo Hospital Surabaya. Result: Most patients with TBM were women (54.29%). Based on age groups, most dominant was adult population (65.71%). Proportion percentage of positive M. tuberculosis complex among 180 specimens were 19.44%. First line anti TB drug sensitivity pattern of 35 isolates were 1 monoresistant, 1 poly-resistant, no multiple drug resistant (MDR), and 33 pan-susceptible. Conclusion: Positivity rate of Mycobacterium tuberculosis complex laboratory diagnosis from TBM suspect patients were low. There was no MDR TB in this study, but mono-resistant and poly-resistant. Microbiological diagnosis was important to give information of active disease and drug sensitivity pattern. Resistance to first line anti TB drugs is alarming to properly manage TBM patients.

scholarly journals RAPID AND SPESIFIC DETECTION OF MYCOBACTERIUM TUBERCULOSIS USING POLYMERASE CHAIN REACTION

2019 ◽  
Vol 3 (2) ◽  
pp. 83
Author(s):  
Anita Kurniati ◽  
Desak Nyoman Surya Suameitra Dewi ◽  
Ni Nyoman Purwani
Keyword(s):  
Polymerase Chain Reaction ◽  
Mycobacterium Tuberculosis ◽  
Clinical Specimen ◽  
Laboratory Diagnosis ◽  
Nucleic Acid Amplification ◽  
Valuable Insight ◽  
Chain Reaction ◽  
Severe Condition ◽  
Middle Income ◽  
Polymerase Chain

Background: Tuberculosis (TB) is one of the major causes of health burden worldwide, especially in lower middle-income countries. TB is caused by Mycobacterium tuberculosis (MTB) and characterized by severe condition incuding coughing and fever. Purpose: To review the current methods for detection of TB using Polymerase Chain Reaction (PCR). Review: several studies have been done to give valuable insight into TB transmission, diagnosis, and treatment, however research  is constantly  needed  to decrease the incidence of eradicate TB. This infectious disease still give big health problem in all over the world by being second in causing high mortality rates after HIV/AIDS.  A specific, sensitive, rapid and cheap method for TB and other mycobacteria diagnosis in clinical specimen is a desperate needed in the laboratory diagnosis and hence management of tuberculosis. PCR as one of nucleic acid amplification assays have revolutionized MTB detection. Since it was first invented in fifteen years ago, it’s been through many developments. Conclusion: PCR  is one of the most specific and sensitive method currently available for TB diagnosis that can also detect in in all types of specimens obtained from TB patients.

scholarly journals Detection of Mycobacterium tuberculosis complex organisms in the stools of patients with pulmonary tuberculosis

Microbiology ◽  
2009 ◽  
Vol 155 (7) ◽  
pp. 2384-2389 ◽  
Author(s):  
Amel El Khéchine ◽  
Mireille Henry ◽  
Didier Raoult ◽  
Michel Drancourt
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Pulmonary Tuberculosis ◽  
Respiratory Tract ◽  
Real Time ◽  
Microscopic Examination ◽  
Real Time Pcr ◽  
Correct Diagnosis ◽  
Signs And Symptoms ◽  
Laboratory Diagnosis ◽  
Acid Fast Bacilli

The laboratory diagnosis of pulmonary tuberculosis mainly relies on the detection of Mycobacterium tuberculosis complex (MTC) organisms in the sputum. In patients who do not give sputum, alternative respiratory tract specimens can be obtained only by invasive procedures. Based on the known survival of MTC organisms in the gastric fluid, we hypothesized that swallowed MTC organisms would be detectable in stool samples. We compared the presence of MTC organisms in respiratory tract specimens and stool specimens collected in parallel from the same patients. MTC was detected in cultures grown on egg-based medium after appropriate decontamination, by microscopic examination after Ziehl–Neelsen staining and by real-time PCR detection of IS6110 using internal controls. A case of pulmonary tuberculosis was defined by the presence of (i) clinical and radiological signs and symptoms suggestive of pulmonary tuberculosis, and (ii) culture of MTC organisms from at least one respiratory tract specimen or (iii) the presence of acid-fast bacilli in the sputum that were subsequently identified as MTC organisms by real-time PCR. The observation of 134 patients suspected to be suffering pulmonary tuberculosis led to the identification of 24 cases and 110 non-infected control patients. Cases and controls did not significantly differ with respect to sex but cases were significantly younger than controls. The sensitivity/specificity was 37.5 %/100 % for the microscopic examination of stools, 54.2 %/100 % for culturing and 100 %/97.3 % for real-time PCR. The positive predicted value was 100 %, 100 % and 88.9 %, respectively, and the negative predicted value was 88 %, 90.9 % and 100 %, respectively. In four patients, a stool specimen initially yielded the correct diagnosis of pulmonary tuberculosis before evaluation of the respiratory tract specimen confirmed the diagnosis. These data indicate that stools could be used in conjunction with sputum testing or as an alternative specimen upon which to base the diagnosis of pulmonary tuberculosis by molecular identification of acid-fast bacilli and culture. This non-invasive alternative procedure is of particular interest for patients who cannot expectorate.

scholarly journals Laboratory Diagnosis of Mycobacterium tuberculosis Infection and Disease in Children

2016 ◽  
Vol 54 (6) ◽  
pp. 1434-1441 ◽  
Author(s):  
James J. Dunn ◽  
Jeffrey R. Starke ◽  
Paula A. Revell
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Latent Infection ◽  
Laboratory Diagnosis ◽  
Molecular Tools ◽  
Mycobacterium Tuberculosis Infection ◽  
Content Type ◽  
Advanced Technologies ◽  
Tuberculosis In Children ◽  
Immunodiagnostic Tests ◽  
Tuberculosis Disease

Diagnosis of tuberculosis in children is challenging; even with advanced technologies, the diagnosis is often difficult to confirm microbiologically in part due to the paucibacillary nature of the disease. Clinical diagnosis lacks standardization, and traditional and molecular microbiologic methods lack sensitivity, particularly in children. Immunodiagnostic tests may improve sensitivity, but these tests cannot distinguish tuberculosis disease from latent infection and some lack specificity. While molecular tools like Xpert MTB/RIF have advanced our ability to detectMycobacterium tuberculosisand to determine antimicrobial resistance, decades old technologies remain the standard in most locales. Today, the battle against this ancient disease still poses one of the primary diagnostic challenges in pediatric laboratory medicine.

scholarly journals Rapid and Visual Differentiation of Mycobacterium tuberculosis From the Mycobacterium tuberculosis Complex Using Multiplex Loop-Mediated Isothermal Amplification Coupled With a Nanoparticle-Based Lateral Flow Biosensor

2021 ◽  
Vol 12 ◽  
Author(s):  
Xinggui Yang ◽  
Junfei Huang ◽  
Xu Chen ◽  
Ziyu Xiao ◽  
Xiaojuan Wang ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Limit Of Detection ◽  
Clinical Specimen ◽  
Lamp Assay ◽  
Lateral Flow ◽  
Isothermal Amplification ◽  
Loop Mediated Isothermal Amplification ◽  
Whole Process ◽  
Specimen Processing ◽  
Mycobacterium Africanum

Tuberculosis (TB) is a chronic infectious disease mainly caused by Mycobacterium tuberculosis (MTB), but other members of the Mycobacterium tuberculosis complex (MTBC), especially Mycobacterium bovis (pyrazinamide-resistant organisms), may also be involved. Thus, the ability to rapidly detect and identify MTB from other MTBC members (e.g., M. bovis, Mycobacterium microti, Mycobacterium africanum) is essential for the prevention and treatment of TB. A novel diagnostic method for the rapid detection and differentiation of MTB, which employs multiplex loop-mediated isothermal amplification (mLAMP) combined with a nanoparticle-based lateral flow biosensor (LFB), was established (mLAMP-LFB). Two sets of specific primers that target the IS6110 and mtp40 genes were designed according to the principle of LAMP. Various pathogens were used to optimize and evaluate the mLAMP-LFB assay. The optimal conditions for mLAMP-LFB were determined to be 66°C and 40 min, and the amplicons were directly verified by observing the test lines on the biosensor. The LAMP assay limit of detection (LoD) was 125 fg per vessel for the pure genomic DNA of MTB and 4.8 × 103 CFU/ml for the sputum samples, and the analytical specificity was 100%. In addition, the whole process, including the clinical specimen processing (35 min), isothermal amplification (40 min), and result confirmation (1–2 min), could be completed in approximately 80 min. Thus, mLAMP-LFB is a rapid, reliable, and sensitive method that is able to detect representative members of MTBC and simultaneously differentiate MTB from other MTBC members, and it can be used as a potential screening tool for TB in clinical, field, and basic laboratory settings.

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