Our Vison ti future on diagnostic Microbiology by Dr.T.V.Rao MD


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CHALLENGES IN DIAGNOSTIC MICROBIOLOGY (Overcoming with Newer Approaches in Diagnostic Microbiology) by Dr.T.V.Rao MD


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CHALLENGES IN DIAGNOSTIC MICROBIOLOGY Overcoming with Newer Approaches in Diagnostic Microbiology Dr.T.V.Rao MD When we are Young in the Microbiology departments the matters were simple with many traditional approaches in diagnosis of Bacterial Parasitic and Fungal infections solving the problems Today the Diagnostic Microbiology is ignited with onset of AIDS as many common uncommon and rare Microbes starting engulfing the humans Profession started to look ahead to the future. In the twenty-first century the clinical microbiology laboratory plays a central part in optimizing the management of infectious diseases and surveying local and global epidemiology. This pivotal role is made possible by the adoption of rational sampling point-of-care tests extended automation and other new technologies including mass spectrometry for colony identification real-time genomics for isolate characterization and versatile and permissive culture systems. However many of our diagnostic laboratories are in primitive stage However we have a larger responsibility to improve the Human care with optimal Diagnostic Microbiology INFECTIONS AND ENVIRONMENT - The emergence of new infections and spread dependent on migration of Humans to Urban in search of employment and livelihood It would seem that any urbanisation drives up the emergence of infectious diseases in those areas Climate change affects infectious diseases directly and indirectly. Meteorological factors including temperature precipitation humidity and radiation influence infectious disease by modulating pathogen host and transmission pathways. Meteorological disasters such as droughts and floods directly impact the outbreak and transmission of infectious diseases. Climate change indirectly impacts infectious diseases by altering the ecological system including its underlying surface and vegetation distribution. EMERGING CHALLENGES TO MEDICAL MICROBIOLOGISTS What the Hospitals and Administrators expect from us - Today’s Microbiology labs are facing growing workloads and the need to produce results faster than ever before. Labs are looking for new ways to optimize lab processes and provide quicker results However Conventional Methods Continues to be main work Since the 1960s numerous ingenious innovations have been introduced and used in clinical microbiology laboratories. Parenthetically none of them allowed microbiologists to abandon the dogma of the pure culture techniques. Never forget Laboratory Medicine Under threat Microbiology is no exception- Great questions to many Microbiologists. Problems with Traditional Methods Cultivation-based methods insensitive for detecting some organisms. Cultivation-based methods limited to pathogens with known growth requirements. Poor discrimination between microbes with common behavioural features. Failure to detect infections caused by uncultivated e.g. novel organisms or organisms that fail to elicit a detectable host immune response. Visual appearance of microorganisms is nonspecific Examples of Failures with Traditional Approaches

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Detection and speciation of slow-growing organisms takes weeks e.g. M. tuberculosis. A number of visible microorganisms cannot be cultivated e.g. Whipple bacillus. Diseases presumed to be infectious remain ill-defined with not detected microorganism e.g. abrupt fever after tick bite. NEED FOR AUTOMATION IN INFECTIOUS DISEASES - Microbes are ever evolving with man we are on the cusp of a dramatic change that will sweep a wave of automation into clinical microbiology laboratories. Threats faced by ecological changes and ever growing misuse of antibiotics for trivial conditions NEED FOR GENOMIC METHODS The initiation of new molecular technologies in genomics and proteomics is shifting traditional techniques for bacterial classification identification and characterization in the 21st century toward methods based on the elucidation of specific gene sequences or molecular components of a cell. Genotypic methods of microbe identification include the use of: Nucleic acid probes PCR RT-PCR RAPD-PCR Nucleic acid sequence analysis 16s rRNA analysis RFLP RAPID IDENTIFICATION SAVES LIVES a number of other studies confirm the urgency of rapid identification of pathogens and its benefit to survival and costs and the effective targeting of antimicrobial therapy The Enigma of Tuberculosis Nearly a third of the worlds population is estimated to be infected with Mycobacterium tuberculosis. This enormous pool of latently infected individuals poses a major hurdle for global tuberculosis TB control. Currently diagnosis of latent TB infection LTBI relies on the tuberculin skin test TST a century-old test with known limitations. Smear examination for Acid fast bacilli continues to be Gold standard with many limitations. Limitations of RNTCP Program However the word TB Continues to be least explored disease the treatment in RNTCP as only National programme to control tuberculosis to be challenged by many clinicians who take care of patients however on many fronts the physicians continue to have their own choices and on many fronts empirical treatment continues and the researcher continue to explore MDR tuberculosis as a threat to the many in the society. Research on new TB diagnostic tools has been accelerated over the last few years and the diagnostic pipeline has been growing rapidly as a result Xpert MTB/RIF Xpert MTB/RIF is an automated cartridge-based nucleic amplification assay for the simultaneous detection of TB and rifampicin resistance directly from sputum in under two hours. The technology is based on the Gene pert platform and was developed as a partnership between the Foundations for Innovative New Diagnostics FIND the test simultaneously detects TB and rifampicin drug resistance a reliable indicator for MDR TB in sputum.

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Emerging Pathogens: Challenges and Success of Molecular Diagnostics - More than 50 emerging and re-emerging pathogens have been identified during the last 40 years. The feasibility of applying molecular diagnostics to dangerous fastidious and uncultivated agents for which conventional tests do not yield timely diagnoses has achieved proof of concept for many agents but widespread use of Molecular Assays. MOLECULAR METHODS IN DIAGNOSIS - The introduction of molecular methods will not only depend on their performance for each individual microorganism but also on the clinical relevance of the diagnostic question asked the prevalence of the clinical problem and whether the new methods are added to the procedures in use or will replace them. Therefore no general rules can be proposed strategies have to be elaborated for each infectious agent or clinical syndrome. DIAGNOSTIC MICROBIOLOGY CHANGING FROM PHENOTYPIC METHODS TO MOLECULAR METHODS In hospital epidemiology the use of such techniques has already provided tests with exceptional discriminatory power. Molecular techniques allow more efficient typing of all pathogens and permit discrimination between strains of organisms that were previously phenotypically identical or uncharacterizable. Currently cost and complexity limit the applicability of these techniques however they are likely to be developed for routine laboratory use in the next decade and their impact will be considerable. METHODS WE SHOULD LOOK TO IMPROVE THE DIAGNOSTIC MICROBIOLOGY 1 BacT/ALERT® Culture Media 2 VITEK® MS 3 MALDI-TOF MS REAL TIME PCR Real-time PCR has established itself as a sensitive and specific qualitative and quantitative technique that has become important to all areas of microbiology. The use of real-time PCR in microbial diagnostics has increased to the point where it has evolved from a novelty into a mature and essential technology for the field. In doing so real-time PCR has driven significant changes in the way we detect microbes. Real-time PCR has engendered wider acceptance of the PCR technique due to its improved rapidity sensitivity reproducibility and the considerably reduced risk of carry-over contamination. Nucleic acid probes Nucleic acid hybridization is one of the most powerful tools available for microbe identification Hybridization detects for a specific DNA sequence associated with an organism. The process uses a nucleic acid probe which is specific for that particular organism. The target DNA from the organism is attached to a solid matrix such as a nylon or nitrocellulose membrane. Tele Diagnosis in Parasitology The CDC now offers tele diagnosis to help laboratories diagnose malaria and other parasitic diseases. When laboratories are not certain about identifying parasites on a slide they can e-mail to the CDC images of the suspected parasites. Experts then review the images and discuss findings with the

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submitting lab within only a few hours allowing near real-time diagnosis as well as an opportunity for training in microscopic diagnosis. Robotics Entering Diagnostic Laboratories Advances to come include programmable automated work stations that can perform a multitude of tasks extremely sophisticated robotics that can perform tasks once restricted to humans and artificial intelligence systems on chips that can learn from their experiences much like ordinary humans. TIME TO MOVE FROM PRACTICE TO GOOD PRACTICES We are moving from just doing the basic work in Diagnostic Microbiology to opting for Best practices as every division of Medicine moving from just attending and treating the patients to practice best practices It is a new trend in Medicine However the Practicing Microbiologists should STRIVE TO IMPROVE DIAGNOSTIC MICROBIOLOGY for BEST PRACTICES. Planning is an essential step in any exercise to rank the risk of communicable disease threats. Establishing the objectives of the exercise enables the selection of an appropriate methodology that is fit for purpose NEVER FORGET LABORATORY REPORTING IS THE MIRROR OF strengths and weaknesses - Reporting results means most of the process can be understood and assessed by multiple stakeholders particularly our physicians and surgeons. Furthermore it enables others to replicate develop and improve upon previous practice leading to improvements in methodologies Ref 1Emerging infectious diseases and cities By James Byrne Scientific American Ref 2Emerging challenges in Infectious Diseases CDC resources Dr.T.V.Rao MD Professor of Microbiology Email doctortvraogmail.com The Document made for improving our vision to Diagnostic Microbiology for the changing needs in Infectious Diseases Available as open resource on www.slideshare.com www.scribd.com www.likeden.com www.autourstream.com Google+ searches

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