Molecular detection of drugresistant tuberculosis by line. A commercial line probe assay for the rapid detection of. Sample dna is hybridized to probes on a membrane strip. The following steps are for the development of the assays manually. In this study, the strategy of testing sputum with genotype mtbdrplus mtbdrplus and. Represents only deaths due to tb disease or tb drug. Aid tb resistance line probe assay for rapid detection of resistant mycobacterium tuberculosis in clinical samples. The lowest concentration of an anti tb agent in vitro that will inhibit the growth of 99% of phenotypically wild type strains of mycobacterium tuberculosis complex mtbc. J global infect dis serial online 2015 cited 2020 may 3.
We would like to show you a description here but the site wont allow us. Immunodiagnostic tests may improve sensitivity, but. The genotype mtbdrplus line probe assay is a suitable and applicable method for establishing the rapidness in detection of drugresistant tb in clinical laboratory. Susceptibility testing using phenotypic methods is labour intensive and timeconsuming. Line probe assay connecting our members with evidence.
Line probe assays lpas enable rapid drugsusceptibility testing for rifampicin rif and isoniazid inh resistance and mycobacterium tuberculosis detection. Drug susceptibility testing, line probe assay, multidrug resistant, m. This follows the 2008 recommendation by who on the use of lpas for the rapid detection of rifampicin resistance. During 2008, there were an estimated 440,000 cases of multidrugresistant tb mdr tb, or 3. In such instances, if there is a high index of suspicion for the presence of mtbc, then an alternative, more sensitive method of molecular identification should be used, such as pcr. Molecular line probe assay for the detection of resistance to second line anti tb drugs sllpa diagnostics background multidrug resistant tuberculosis mdr tb is a public health crisis and a global health security risk carrying grave consequences for those affected. Line probe assay has been shown to be highly sensitive and specific in the detection of rifampicinresistant tb when used on culture isolates. Various diagnostic methods, including acid fast stain afs, tb culture, nucleic acid hybridization probes, line probe assays, and nucleic acid amplification tests. Limited performance data from line probe assays lpas, nucleic acid tests used for the rapid diagnosis of tuberculosis tb, nontuberculosis mycobacteria ntm, and mycobacterium tuberculosis drug resistance are available for hivinfected individuals, in whom paucibacillary tb is common. Our findings supports the use of the mtbdr line probe assay for rapid m.
Infections caused by mycobacterium tuberculosis complex remain one of the most important global public health issues. Most likely it is the only firstline drug to be maintained in all future tb regimens. Handbook on tb laboratory diagnostic methods in the. Tb assay requires four wells to be used for each patient sample. Tb lipa is a commercial line probe assay designed to rapidly detect. Who recommends this rapid diagnostic test for identifying those. Costeffectiveness of a line probe assay test compared to. Technical faqs on hain genotype mtbdrplus line probe assay 2 conventional tests for tb involve afb smear, culture and dst. Comparison of line probe assay to bactec mgit 960 system. Gupta r, thakur r, gupta p, jalan n, kushwaha s, gupta m, gupta p, aggarwal a, manchanda v.
The mtbdrplus and mtbdrsl line probe assays were evaluated in 181 sputum samples and 270 isolates from patients with cultureconfirmed drugsensitive tb, mdr tb, or xdr tb. The line probe assay is used for the diagnosis of mdr tb. Rapid detection of multi drug resistant tuberculosis using. Pdf line probe assay for detection of mycobacterium tuberculosis.
Pdf comparison of xpert mtbrif with line probe assay. The recommended use of line probe assays is currently limited to m. Tb resistant to first line drugs but also have the potential to become an important part of new tb regimens 4. Use and evaluation of a line probe assay in patients with. Correlation between genotypic and phenotypic testing for. Recently, als assay nods the scientific community as it is rapidly used for diagnosis of tuberculosis. Tuberculosis most common infectious cause of mortality after hivaids 2 3. Line probe assay connecting our members with evidence and. Role of molecular methods in tuberculosis diagnosis and. Laboratory diagnosis of mycobacterium tuberculosis. Npt tee customer supplied pps threaded adaptor flow flexible conduit and coupling customer supplied tbx551. The performance of the lineprobe assays relative to culturebased ds tests was evaluated in metaanalyses 35.
Erln tb european reference laboratory network for tuberculosis 20102014 erltbnet european reference laboratory network for tuberculosis from 2014. Both tests are based on detection of mutations in the rifampin rif resistancedetermining region of dnadependent rna polymerase gene rpob. The target dna, previously extracted by boiling, is pcramplified using biotinylated primers and finally incubated with the strip. Accuracy of line probe assays for the diagnosis of. February 20 this report contains the collective views of an international group of experts, and does not necessarily represent the decisions or the stated policy of the world health organization. Pdf evaluation of genexpert mtbrif and line probe assay. What sample was used to diagnose the tb and what was the reading on the bands. It is an infectious disease caused by the bacillus mycobacterium tuberculosis.
This platform uses a nitrocellulose strip for identification of mtbc and commonly isolated ntm by reverse hybridization. Known positive and negative control strains were tested for rapid tb antigen per new batch 9, 10, 11. The probe is used as an additional test when there is not a clear id off the high performance liquid chromatography hplc mycolic acid analysis. Line probe assay for detection of mycobacterium tuberculosis. Conventional dst is carried out on solid or, more recently, in liquid media after detection and identification of the pathogen previously grown from clinical specimens.
The genotype mtbdrplus test is a whoendorsed lineprobe assay lpa for the. Tb online who recommends use of two alternative line. At that time, the evidence assessed was for the lpa technology, hain version 1 assay. The conventional tuberculosis drug susceptibility tests are sensitive and specific, but they are not rapid. The deoxyribonucleic acid dna probe procedure is used to identify the mycobacteria species of mycobacterium tuberculosis complex mtbc and m. Appendix 1 rapid molecular testing laboratory info tb cdc. Accuracy of line probe assays for the diagnosis of pulmonary and multidrugresistant tuberculosis. The mtbdrplus line probe assay lpa and xpert mtbrif have been endorsed by the world health organization for the rapid diagnosis of drugresistant tuberculosis. Comparative evaluation of genotype mtbdrplus line probe assay. The line probe assays detect drugresistant tuberculosis in clinical samples. Line probe assay conclusion advantages easy to perform detection of 18 clinically relevant mycobacteriumsp. Lineprobe assay and molecular typing reveal a potential. Gli model tb diagnostic algorithms stop tb partnership. In multidrug resistant tuberculosis mdrtb infectiousness is frequently prolonged, jeopardizing efforts to control tb.
The line probe assay was able to diagnose mtbc in 38. While afbnegative sputum decreases test sensitivity, use in this setting may still be considered, because mtbdr. Appendix 1 rapid molecular testing laboratory info. However, until this study there have been few data on direct testing of. However, there is no clarity regarding the superiority of one over the other. Elagib rapid detection of multi drug resistant tuberculosis using line probe assay lpa in sudan european academic research vol. Diagnosis of tuberculosis in children is challenging.
Introduction tuberculosis tb is the leading cause of death in the world. The world health organization has recommended use of molecularbased tests mtbdrplus and genexpert mtbrif to diagnose multidrugresistant tuberculosis in developing and highburden countries. Susceptibility testing using phenotypic methods is labour intensive and time. New tb diagnostic tools and the challenges of interpreting. Muhammad yahya noori assistant professor department of pathology dimc focal person for provincial tuberculosis reference lab sindh duhs 2. This assay became the gold standard for tb drug susceptibility testing dst. Comparison of xpert mtbrif with line probe assay for. Tb innogenetics, zwijndrecht, belgium, a commercially available kit not yet approved by the u. Test mtd, gen probe xpert mtbrif often called genexpert p i psqpyrosequencing psq home brews the genthe gen probe mtd testprobe mtd test the first fda approved naat for m. Such mutations are found in 9598% of mycobacterium. Evaluation of geno type mtbdrplus line probe assay for early detection of drug resistance in tuberculous meningitis patients in india. Antibodies from lymphocyte secretion or antibody in lymphocyte supernatant or als assay is an immunological assay to detect active diseases like tuberculosis, cholera, typhoid etc. Tbx587, tbx557 or tbx564 ph sensor typical twistlock sensor installation tbx551 ph sensor customer supplied full port ball valve compression fitting process pipevessel 12 to 25 mm 0. The performance of the line probe assays relative to culturebased ds tests was evaluated in metaanalyses 35.
Controlling resistance, reducing transmission and improving treatment outcomes in mdrxdr tb patients is reliant on susceptibility testing. As it is we are not able to ascertain what kind of results you have since lpa will also give inh and rif sensitivity patterns. Division of vir national center for infectious diseases. Dna probe mycobacteria identification state public health. Development of a high sensitivity taqmanbased pcr assay. With the ability to detect lowlevel isoniazid resistance inh a, the whoendorsed genotype mtbdr plus hain lifescience, germany version 1 is an improvement over the mtbdr hain lifescience, germany assay which had only rpob probes to determine rifampicin. The arms are chosen to be complementary to each other, so that under assay conditions they hybridize to form a stemandloop secondary structure in which the probe sequence is located in the loop. Genotype mtbdrsl line probe assay shortens time to. Comparative evaluation of genotype mtbdrplus line probe. Comparison of the mgit tbc immunochromatographic assay with. Molecular assays in tuberculosis jatin yegurla junior resident 1732018.
Methods in this crosssectional study, 269 previously treated sputum. Jul 28, 2005 mycobacterium tuberculosis is a leading cause of death worldwide. Evaluation of geno type mtbdrplus line probe assay for early. Viral hepatitis, std and tb prevention division of viral hepatitis hepatitis reference laboratory. Comparison of xpert mtbrif with line probe assay for detection of rifampinmonoresistant mycobacterium tuberculosis. Background the objectives of the study were to compare the performance of line probe assay genotype mtbdrplus with solid culture method for an early diagnosis of multidrug resistant tuberculosis mdr tb, and to study the mutation patterns associated with rpob, katg and inha genes at a tertiary care centre in north india. Policy guidance on the molecular secondline lineprobe assay pdf. Molecular line probe assay for the detection of resistance to second line anti tb drugs sllpa tuberculosis diagnostics background multidrug resistant tuberculosis mdr tb is a public health crisis and a global health security risk carrying grave consequences for those affected. In a doubleblinded prospective study, we evaluated the efficacy of the xpert mtbrif on samples that were first tested by lpa under the revised national. Another identification option the line probe assay. Line probe assays for drug resistant tuberculosis detection. Technical faqs on hain genotype mtbdrplus line probe assay. May 27, 2014 this study was conducted to assess the efficacy of line probe assay in detecting mtb from smear negative culture negative samples in a tertiary care centre catering to the need of a high burden tb population of uttarakhand.
Tb lamp test eiken chemical, tokyo japan, and line probe assays lpas, have become available to replace or complement existing conventional tests for detecting mycobacterium tuberculosis complex bacteria mtb and for detecting drug resistance. Jan 04, 2016 the line probe assay uses the reverse hybridization technology. Only 25% of multidrugresistant tuberculosis mdr tb cases are currently diagnosed. Pdf accuracy of line probe assays for the diagnosis of. There are three commercially available line probe assays, two by hain and one by innogenetics. Technical faqs on hain genotype mtbdrplus line probe assay 3 is 98. The incidence of multidrugresistant tuberculosis mdr tb is increasing and the emergence of extensively drugresistant tuberculosis xdr tb is a major challenge. Molecular line probe assay for the detection of resistance to isoniazid and rifampicin lpa tuberculosis diagnostics background multidrugresistant tuberculosis mdr tb is a public health crisis and a major threat to global tb control. Handbook on tb laboratory diagnostic methods in the european. Published studies have shown that the assay performs well when using culture isolates. A nil control and a cell functionality positive control should be run with each individual sample. During this period, the world health organization who had issued 10 policy statements for improving diagnosis of tb, including the use of commercial and noncommercial dst methods and implementation of molecular methods such as the line probe assay lpa and xpert mtbrif or genexpert assay. Dna was extracted from the appropriate sample using chemical a and b.
Line probe assay lpa test detects tb infection presence and resistance to two drugs rifampicin and isoniazid. Evaluation of a novel line probe assay to detect resistance. Lpa line probe assay ltbi latent tb infection mdr multidrug resistant miru mycobnereritacpi st esaedl r. The rapid tb anigen sd bioline was tested to confirm the presence of organisms belonging to m. Objective to evaluate the performance of genotype mtbdrplus ver 2. Lpa line probe assay ltbi latent tb infection mdr tb multidrug resistant tuberculosis mg milligram mri magnetic resonance imaging ngo nongovernmental organisation ntp national tb programme ppe personal protective equipment ptb pulmonary tb rr tb rifampicinresistant tb sllpa second line line probe assay tb tuberculosis. Evaluation of two line probe assays for rapid detection of.
Evaluation of a novel line probe assay to detect resistance to. Aid tb resistance line probe assay for rapid detection of. The development of rapid molecular diagnostic assays for pyrazinamide pza resistance is considered technically challenging as mutations are highly diverse, scattered along the full length of the pnca gene and not all are associated with pza resistance. Mycobacterium tuberculosis complex drug susceptibility. There is a paucity of data on application of this test directly to clinical specimens, although based on a small number of studies, the test. Dec 28, 2017 the incidence of multidrugresistant tuberculosis mdr tb is increasing and the emergence of extensively drugresistant tuberculosis xdr tb is a major challenge. We evaluated the performance of the novel genoscholar pza tb ii line probe assay pzalpa2.
Clinical diagnosis lacks standardization, and traditional and molecular microbiologic methods lack sensitivity, particularly in children. Line probe assays lpas enable rapid drugsusceptibility testing for rifampicin rif and isoniazid inh. The diagnostic utility of line probe assays for multidrug. Evaluation of a novel line probe assay to detect resistance to pyrazinamide, a key drug. Evaluation of a novel line probe assay to detect resistance to pyrazinamide, a key drug used for tuberculosis treatment. Line probe assay all 161 selected isolates were heatinactivated and tested by genotype mtbdrplus v1. Every assay is optimized to run under universal thermal cycling conditions with a final reaction concentration of 250 nm for the probe and 900 nm for each primer. Can use mgit liquid culture and solid media quick visual interpretation mostly automated using the auto. Genotype mtbdrplusv1 was whoendorsed in 2008 but newer lpas have since been developed.
Methods a total of 572 ziehlneelsen sputum smearnegative samples were selected. Pdf utility of line probe assay for the early detection of. Tb online who recommends use of two alternative line probe. A liquid culture will be the most definitive way of ruling out tb. A lab does the test and its responsibility of a physician to read and inform the result to a non medical patient or relative who is undergoing diagnostic test. Diagnostic utility of a line probe assay for multidrug resistanttb in. If the clinical picture is highly suggestive of tb, a repeat test can be ordered to increase sensitivity. Microscopy cannot detect drug resistance, while the mtbdrplus can rapidly detect resistance to rifampicin and isoniazid, the most important first line tb drugs. The mtbdrsl test is a molecular line probe assay for the rapid detection of mutations associated with resistance to fluoroquinolone and aminoglycoside antibiotics and cycloserine, drugs used to treat multidrugresistant tuberculosis. Comparative analysis of line probe assay and conventional. Evaluation of genexpert mtbrif and line probe assay for rapid diagnosis of mycobacterium tuberculosis in sudanese pulmonary tb patients. Another study comparing the bd tbc antigen assay with the gen probe tb assay showed similar discrepant results said et al.
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