Newer diagnostic methods in tuberculosis detectionApollo Hospitals
One-third of the world's population has been infected with Mycobacterium tuberculosis, with new infections occurring in about 1% of the population each year. However 90–95% of infections remain asymptomatic. Thus early diagnosis of tuberculosis and drug resistance improves survival and helps to promote contact tracing, implementation of institutional cross-infection procedures, and other public-health actions. There have been many advances and modifications to the methodology for tuberculosis diagnosis some of which are very promising. But these advances have not kept pace with the explosion of tuberculosis or the outbreak of drug resistant tuberculosis. This review describes some of the newer advances in tuberculosis diagnostics and the challenges they face.
This presentation is about lab diagnosis of tuberculosis. It highlights use of currently available diagnostic methods in identifying pulmonary and extrapulmonary tuberculosis.
Newer diagnostic methods in tuberculosis detectionApollo Hospitals
One-third of the world's population has been infected with Mycobacterium tuberculosis, with new infections occurring in about 1% of the population each year. However 90–95% of infections remain asymptomatic. Thus early diagnosis of tuberculosis and drug resistance improves survival and helps to promote contact tracing, implementation of institutional cross-infection procedures, and other public-health actions. There have been many advances and modifications to the methodology for tuberculosis diagnosis some of which are very promising. But these advances have not kept pace with the explosion of tuberculosis or the outbreak of drug resistant tuberculosis. This review describes some of the newer advances in tuberculosis diagnostics and the challenges they face.
This presentation is about lab diagnosis of tuberculosis. It highlights use of currently available diagnostic methods in identifying pulmonary and extrapulmonary tuberculosis.
RECENT ADVANCES IN DIAGNOSIS OF TUBERCULOSISANGAN KARMAKAR
TRADITIONAL TESTS AND RECENT DIAGNOSTIC MODALITIES FOR TUBERCULOSIS WITH EMPHASIS TO MOLECULAR DETECTION TECHNIQUES, DRUG SENSITIVITY ASSESMENT IN INDIAN PERSPECTIVE
Information about diagnostic methods and techniques for tuberculosis including microscopy, fluorescence microscopy, mycobacterial culture, molecular techniques (line probe assay, Xpert MTB/RIF), interferon gamma release assay (IGRA) and tuberculin skin test (TST)
RECENT ADVANCES IN DIAGNOSIS OF TUBERCULOSISANGAN KARMAKAR
TRADITIONAL TESTS AND RECENT DIAGNOSTIC MODALITIES FOR TUBERCULOSIS WITH EMPHASIS TO MOLECULAR DETECTION TECHNIQUES, DRUG SENSITIVITY ASSESMENT IN INDIAN PERSPECTIVE
Information about diagnostic methods and techniques for tuberculosis including microscopy, fluorescence microscopy, mycobacterial culture, molecular techniques (line probe assay, Xpert MTB/RIF), interferon gamma release assay (IGRA) and tuberculin skin test (TST)
The following presentation is only for quick reference. I would advise you to read the theoretical aspects of the respective topic and then use this presentation for your last minute revision. I hope it helps you..!!
Mayur D. Chauhan
ABSTRACT- Multiple Drug resistance (MDR) tuberculosis timely diagnose is of utmost clinical relevance and needs to be diagnose at initial stages for the proper treatment. The current study was done to detect the several genes for MDR tuberculosis (TB) in clinical isolates by molecular tools. 60 clinical isolates were collected and subjected for AFB smear preparation, Nested PCR (IS6110) for mycobacterium tuberculosis complex detection and MDR TB PCR targeting rpoB, kat G, mab A promoter. 12 came positive for AFB smears, out of which 08 were pulmonary and 04 were extra pulmonary. Nested PCR targeting IS6110 gene was amplified at 123 base pairs with 340 base pairs as IC (internal control) was seen in 25 cases which include 19 pulmonary and 6 extra pulmonary. The Positive TB PCR specimens were subjected for MDRTB PCR Only 06 cases yielded, an amplicon of 315 bp confirming the rpoB gene resistance for resistance for rifampcin drug. In any of the 06 positives none of the other resistance gene other than rpoB was amplified. Targeting multiple genes at once, additional information will be gained from a single test run that otherwise would require several times the reagents and more time to perform. Current study signifies the usage of quick, cost effective, DNA sequences based method for MDR TB detection where disease will be diagnosed earlier and hence treatment would be started at an early stage.
Keywords: Multiple drug resistance, amplicon, Polymerase chain reaction, Nested PCR, Rifampicin.
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The bacteria that cause tuberculosis (TB) can develop resistance to the antimicrobial drugs used to cure the disease. Multidrug-resistant TB (MDR-TB) is TB that does not respond to at least isoniazid and rifampicin, the 2 most powerful anti-TB drugs.
The 2 reasons why multidrug resistance continues to emerge and spread are mismanagement of TB treatment and person-to-person transmission. Most people with TB are cured by a strictly followed, 6-month drug regimen that is provided to patients with support and supervision. Inappropriate or incorrect use of antimicrobial drugs, or use of ineffective formulations of drugs (such as use of single drugs, poor quality medicines or bad storage conditions), and premature treatment interruption can cause drug resistance, which can then be transmitted, especially in crowded settings such as prisons and hospitals.
In some countries, it is becoming increasingly difficult to treat MDR-TB. Treatment options are limited and expensive, recommended medicines are not always available, and patients experience many adverse effects from the drugs. In some cases even more severe drug-resistant TB may develop. Extensively drug-resistant TB, XDR-TB, is a form of multidrug-resistant TB with additional resistance to more anti-TB drugs that therefore responds to even fewer available medicines. It has been reported in 117 countries worldwide.
Drug resistance can be detected using special laboratory tests which test the bacteria for sensitivity to the drugs or detect resistance patterns. These tests can be molecular in type (such as Xpert MTB/RIF) or else culture-based. Molecular techniques can provide results within hours and have been successfully implemented even in low resource settings.
New WHO recommendations aim to speed up detection and improve treatment outcomes for MDR-TB through use of a novel rapid diagnostic test and a shorter, cheaper treatment regimen. At less than US$ 1000 per patient, the new treatment regimen can be completed in 9–12 months. Not only is it less expensive than current regimens, but it is also expected to improve outcomes and potentially decrease deaths due to better adherence to treatment and reduced loss to follow-up.
Solutions to control drug-resistant TB are to:
cure the TB patient the first time around
provide access to diagnosis
ensure adequate infection control in facilities where patients are treated
ensure the appropriate use of recommended second-line drugs.
In 2015, an estimated 480 000 people worldwide developed MDR-TB, and an additional 100 000 people with rifampicin-resistant TB were also newly eligible for MDR-TB treatment. India, China, and the Russian Federation accounted for 45% of the 580 000 cases. It is estimated that about 9.5% of these cases were XDR-TB.
La disponibilidad de un sistema de multiplicación del virus de la hepatitis C (VHC) infeccioso en cultivos celulares está permitiendo investigar nuevos factores de respuesta a tratamientos antivíricos en condiciones controladas. Se presentará evidencia de que el fitness vírico puede ser un factor de multiresistencia a inhibidores y quese pueden obtener eficientes reducciones de carga viral empleando diseños secuenciales de administración de inhibidores que incluyan ribavirina. Se discutirán posibilidades de aplicación clínica.
Antiretroviral Resistance in HIV-1 Patients at a Tertiary Medical Institute in Saudi Arabia: a Retrospective Study and Analysis.
Journal Club,
Virology Rotation , 1/5/2019
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This Digital Transformation and IT Strategy Toolkit was created by ex-McKinsey, Deloitte and BCG Management Consultants, after more than 5,000 hours of work. It is considered the world's best & most comprehensive Digital Transformation and IT Strategy Toolkit. It includes all the Frameworks, Best Practices & Templates required to successfully undertake the Digital Transformation of your organization and define a robust IT Strategy.
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The world of search engine optimization (SEO) is buzzing with discussions after Google confirmed that around 2,500 leaked internal documents related to its Search feature are indeed authentic. The revelation has sparked significant concerns within the SEO community. The leaked documents were initially reported by SEO experts Rand Fishkin and Mike King, igniting widespread analysis and discourse. For More Info:- https://news.arihantwebtech.com/search-disrupted-googles-leaked-documents-rock-the-seo-world/
Discover the innovative and creative projects that highlight my journey throu...dylandmeas
Discover the innovative and creative projects that highlight my journey through Full Sail University. Below, you’ll find a collection of my work showcasing my skills and expertise in digital marketing, event planning, and media production.
LA HUG - Video Testimonials with Chynna Morgan - June 2024Lital Barkan
Have you ever heard that user-generated content or video testimonials can take your brand to the next level? We will explore how you can effectively use video testimonials to leverage and boost your sales, content strategy, and increase your CRM data.🤯
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PPT Cirillo " Diagnostic laboratory news"
1. Le novità diagnostiche di laboratorio Daniela Maria Cirillo WHO Collaborating Centre for integrated laboratory strengthening on TB San Raffaele Scientific Institute Milano 14/15 ottobre 2011
27. Comparison GenoType® MTBDRplus and INNO-LiPA Rif.TB GenoType ® MTBDR INNO-LiPA Rif.TB Company Hain Lifescience Innogenetics M. tuberculosis detection Yes Yes Detection of RMP Resistance in M. tb Complex Yes Yes Detection INH Resistance in M. tb Complex Yes No Strip Assay Yes Yes PCR based Yes Yes From liquid or solid culture Yes Yes Direct assay Yes Yes (modified version) TBC Detection: 23S-rRNA/16S-rRNA Yes Yes RMP-Resistance: rpoB gene Yes Yes INH-Resistance: katG gene/inhA gene Yes No Universal control Yes No rpoB unicontrol Yes No kat G/inHA u nicontrol Yes No
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29. New generation of LiPA performs better in both Sm+ and Sm- samples Miotto al. JCM 2008 TBPANNET workpackage 6 midterm meeting
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31. Molecular line probe assays for rapid screening of patients at risk of MDR-TB Policy statement by WHO and Partners June 27, 2008 Endorsement of the two commercial line probe assays for rifampicin resistance detection TBPANNET workpackage 6 midterm meeting
32. Possible automation on LiPA LiPAs require: Level II biosafety areas Skilled laboratory staff Amplicon Contamination control
51. TBPANNET workpackage 6 midterm meeting PMA pretreatment of samples allows selective amplification of DNA from live bacteria Miotto 2011 under revision ERJ
53. Typing: the gold standard Mycobacterial Interspersed Repetitive Units -Variable Number of Tandem Repeats (MIRU-VNTR) Based on tandem repeats in mini-satellite regions of the genome. The original 15 loci consisted of 5 exact tandem repeats (ETR A-E) followed by 10 MIRUs. Described by Supply et al ., 2006. VNTR-424 VNTR-1955 VNTR-2163b VNTR-2347 VNTR-2401 VNTR-3171 VNTR-3690 VNTR-4052 VNTR-4156 VNTR locus MIRU2 MIRU4 (ETR-D) MIRU10 MIRU16 MIRU20 MIRU23 MIRU24 MIRU26 MIRU27 MIRU31 (ETR-E) MIRU39 MIRU40 ETR-A ETR-B ETR-C
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56.
57.
58. 2.) T-cell based tests APC T cell antigens/ peptides cytokine induction cytokine induction cytokine induction ELISA ELISPOT assay T.SPOT. TB QuantiFERON TB gold IGRA IFN- release assay PPD (= tuberculin) ESAT-6, CFP-10, Tb7.7 ESAT-6, CFP-10 Skin test
59. Immunology based Tb-tests Serological tests Antibodies against Mtb-specific antigens Skin tests Local immune reaction (PPD-specific T cells) Interferon gamma release assays (IGRAs) Mtb-specific T cells (PPD, ESAT-6, CFP-10) APC T cell antigens/ peptides
61. Summary of pooled values from the metanalysis performed by Pai et al, and by Sester and Sotgiu et al Test Sensitivity for active TB Specificity for active TB Percentage TST Pai et al, 2008 77 95/79 Sester et Sotgiu et al, 2010 65 75 QFT-IT Pai et al, 2008 70 96 Sester et Sotgiu et al, 2010 80 79 T-SPOT. TB Pai et al, 2008 90 93 Sester et Sotgiu et al, 2010 81 59
62. IGRA in HIV+ with active TB Source Patient number Test Sensitivity (indeterminate results included) Sensitivity (indeterminate results excluded) Seshandri et al, 2008 13 QFT 23 33 Vincenti et al, 2007 13 QFT ND 85 Raby et al, 2008 59 QFT-IT 63 76 Aabye et al, 2009 68 QFT-IT 65 83 Leidl, ERJ 2009 31 QFT-IT 68 68 Kabeer et al, 2009 105 QFT-IT 65 78 Goletti et al, 2010 28 QFT-IT 61 73 Vincenti et al, 2007 13 T SPOT TB ND 85 Markova, 2009 13 T SPOT TB 62 89 Jiang, 2009 32 T SPOT TB 66 66 Leidl, ERJ 2009 19 T SPOT TB 89 100
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64. Sensitivity, specificity and diagnostic odds ratio of the different assays for the immune diagnosis of TB Goletti et al, CMI 2006 Assay Sensitivity % Specificity % Diagnostic odds ratio RD1 ELISPOT assays Selected peptides 70 91 22 Intact proteins 83 56 6 RD1 commercially available assays T-SPOT TB 91 59 15 QFT Gold 83 59 7
66. IGRA at the site of TB disease: BAL vs blood From Jafari, AJRCCM 2009
67. IGRA at the site of TB disease: Pleural fluid vs blood Berlin, October 4th, 2008 From Losi et al, ERJ 2007 PLEURAL CELLS PBMC
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69. Predictive value of IGRA: HIV-negative subjects Diel et al, AJRCCM 2010 in press
70. Rates of disease progression in IGRA+ve vs TST Country Test Incidence of active TB in IGRA+ groups Comment Gambia [Hill et al. 2008] ELISPOT (in-house) 9/1000 person-yr High burden Colombia [del Corral et al. 2009] In-house CFP-10 assay 7/1000 person-yr High Burden Senegal [Lienhardt et al. 2010] ELISPOT (in-house- 32 SFC cut-point) 9/1000 person-yr High burden Turkey [Bakir et al. 2008] ELISPOT (in house similar to T-SPOT TB ) 21/1000 person-yr Intermediate Germany [Diel et al. 2010] QFT-IT 73/ 1000 person-yr Low burden
71. Multifunctional T cells correlate with active TB disease From Caccamo, EIJ 2010, modified IFN- γ , IL-2, TNF- α IFN- γ , IL-2 IFN- γ
Je vous parle d ’ un nouveau test pour le diagnostic de la tuberculose. Pourquoi? Qu ’ est-ce que c ’ est la TB? Et pourquoi a t-on besoin d ’ un nouveau test? Pour répondre a ces questions j ’ ai préparé une liste d ’ objectifs.
A recent meta-analysis assessed the diagnostic accuracy of the commercial NAA tests described above (Ling et al., 2008). Sensitivity (not specificity) of different tests (except real time PCR) depends on samples smear results - positive or negative. It is interesting that NAA tests’ sensitivity of extrapulmonary samples varies much between diffrent tests. Real time PCR shows equal sensitivity for all kind of samples (pulmonary smear positive and negative and extrapulmonary). Specificity of all indicated tests for all kind of samples is nearly the same and quite high. Overall, a pooled sensitivity of 85% (range 36–100%) and specificity of 97%(range 54–100%) was reported. Both values showed significant heterogeneity, which could not be explained even after analysing the results of each test separately. The major conclusion was that the accuracy of these NAATs with respiratory samples was highly variable, with sensitivity giving lower values than specificity. For these reasons, they probably still do not have enough clinical relevance and could not be recommended to replace the conventional tests for diagnosing pulmonary tuberculosis.
ISTC Training Modules 2008 In any given population of TB bacilli, naturally occurring resistant organisms occur at the rates as shown. [Review above] The greater the burden of disease, the larger the bacillary population and the greater the risk for harboring drug-resistant mutants and acquiring drug resistance. [Note: Year drug was discovered is listed under each drug.] [Image Credit: CDC/Dr. Ray Butler; Janice Carr. Illustration Credit: National Institute of Allergy and Infectious Diseases (NIAID). Illustrator: Krista Townsend]
In this slide, MTB/RIF assay procedure is described. In brief, a sputum sample is treated for 15 minutes with sodium hydroxide and isopropanol-containing reagents, which decrease MTB viability by at least 6 logs. The sample is transferred into the cartridge preloaded with PCR buffers. The cartridge is then loaded into the instrument and an automatic process completes the sample processing, DNA extraction and real-time PCR, thus eliminating the risk of cross-contamination. The test turnaround time is less than 2 h, similar to smear microscopy and many time faster than culture methods.
Up to now, the few papers that report the MTB/RIF performance, show for pulmonary specimens, a sensitivity of more than 98% in smear- and cultures positive samples, and of almost 72% in smear negative culture positive samples. The overall specificity is 99-100%. In addition, MTB/RIF correctly detected rifampicine resistance in more than 96% patients, with a high specificity.
Similar sensitivity and specificity percentages are observed when the Xpert MTB RIF performance is evaluated in low MDR- prevalence population. A good performance, even if we see lower values, is observed in extrapulmonary samples and this indication may imply the isolation of TB strains from other sample in particular patients like children.
Finally, here I reported the last WHO recommendations. WHO strongly recommends Xpert MTB/RIF should be used as the initial diagnostic test in individuals suspected of MDR-TB or HIV/TB and it may be used as a follow-on test especially in smear-negative specimens. The expected impact of Xpert MTB/RIF implementation would be a three-fold increase in the diagnosis of patients with MDR-TB and TB/HIV cases and an increase of treated TB at least may reduce mortality, secondary resistance and ongoing transmission.
So, summarizing the Xpert MTB RIF characteristics: It is simple to perform It can be performed by low skilled technicians in absence of biosafety infrastructure and this (next slide) will make Xpert MTB/RIF test suitable for district and sub-district levels. It shows an high sensitivity also in paucibacillary samples simplifying the diagnosis of smear negative TB as HIV+ High performance in extrapulmonary samples and The advantage of MTB and RIF resistance detection within 2 hours However we have no data on Xpert MTB/RIF performance in children and few data in low-prevalence populations It requires uninterrupted and stable electronic power supplies and yearly calibration Size for storage issues
Highlight the high variation between the different commercial kits (e.g. low sensitivity of QFT on extrasanguin but the high sensivity for same sample using T-spot). compare values to that of TST.
Sensitivity of our in house assays based on selected peptides was lower compared to the other assays; however the specificity was much higher and more importantly the diagnostic odds ratio was significantly higher compared to the other tests