Tag: Pyro sequence screening

Abstract Pathogenic organisms are transmitted to the host organism through all possible connectedpathways, and cause severe disease state in the new host. Commonly occurring curable infectious diseases stillimpose the greatest health impacts on a worldwide perspective. Several technologies are used for diagnosing thedisease state in that particular host. Golden standard for diagnostics of pathogenic bacteria has long been cultureable Medias. This method was used from the ancient days to diagnose a disease. Environmental biologists haveestimated that less than 1% of all bacteria are culture able and it takes lots of time to get conformation about thedisease, severity and causative organism. And these are the cost effective techniques. Genomic-based approachesoffer the potential to identify all microbes from all the biological kingdoms. Nucleic acid based pathogendiagnostics has evolved significantly over the past decades. Novel technologies offer increased potential insensitivity, specificity, decreased costs and parallel sample management. However, most methods are confined tocore laboratory facilities. To construct an ultimate nucleic acid based diagnostic for use in areas of need, potentialfrontline techniques need to be identified and combined.The research focus of this work has been to develop and apply nucleic acid based methods for pathogendiagnostics. Methods and assays were applied to the two distinct systems i) screening for antibiotic resistancemutations in the bacterial pathogen Helicobacter pylori appears to be an important risk factor for stomach ulcers,gastritis and possibly stomach cancer and ii) genotype determination of the cancer causative Hepatitis B and Cvirus (HBV & HCV). The first part of the study included development of rapid, direct and multiplex Pyrosequencing nucleic acid screenings. With improved methodology in the sample preparation process, we coulddetect an existence of multiple co-infecting HBV & HCV genotypes at greater sensitivities than previouslydescribed, when using the same type of methodology. The second part of the study focused on multiplex nucleicacid amplification strategies using Molecular Inversion Probes with end-step Pyro sequencing screening. [avf_view name="Article Addons" view-id="66d98451c48e3"]