Sensitivity Of Duplex Real Time Pcr - 1617 Words

Sensitivity of duplex Real-time-PCR compared microscopic detection of M. bovis: Results revealed that out of 600 lymph node sample with lesions suggestive to tuberculosis 580 (96.6%) was positive for AFB detected by microscopic examination of ZN stained smears. However, by duplex real-time PCR 588 (98%) was confirmed to M. bovisinfection. Analytical specificity: The specificity of real-time PCR targeting IS1081 and IS6110 was evaluated to 19 strains of different Mycobacterial species. The real-time PCR targeting both IS1081 and IS6110 sequences showed negative result with all Mycobacterial A. Selim et el. 50 species in two concentrations of DNA from each strain, 5ng/ µl and 5pg/ µl; while strong positive result with M. bovisBCG was detected. Furthermore, ÃŽ ²-actin internal control showed positive Ct-values with all Mycobacterial species including M. bovisBCG (table 1). Table 1. Mycobacteria and non-mycobacteria analyzed for the determination of the specificity of real-time MAP-PCR Target sequence IS1801 IS6110 Template concentration Species, Sub-species Type Host species / Source 5ng/ µl 5pg/ µl 5ng/ µl 5pg/ µl M. avium subspecies avium (M128/2) TS Cattle – – – – (01A1077/2) FI-J Cattle – – – – (00A0720/2) FI-J Pig – – – – (03A0910/2) FI-J Poultry – – – – (03A2530/1) FI-J Poultry – – – – M. avium subspecies hominisuis (01A0554/1) FI-J Pig – – – – (01A1054/1) FI-J Human – – – – (01A0255/1) FI-J Dog – – – – M. bovisBCGShow MoreRelatedEvaluation Of The Method Of Rt Pcr3051 Words   |  13 PagesIntroduction RT PCR is one of the most useful methods used within investigated tissue to compare all transcripts accurately. To apply it accurately it has some challenges as it critically depends on the various reference materials and correct use of calibration. PCR diagnostics application to tissues is often limited by the lack of quality template due to inefficient RNA extraction methodologies (Sellars et al., 2007). Ecologically influenced or disease phenotypes and the cellular expression patternsRead MoreMicrorna Essay1004 Words   |  5 Pagesnucleocytoplasmic shuttle exports pre-miRNA from nucleus to cytoplasm using GTP.[24] Cytoplasmic processing Another enzyme RNase III enzyme Dicer cleaves premiRNA. It cuts away the loop joining the 3’ and 5 ‘arms yielding imperfect miRNA duplex about 22 nucleotides. Functional miRNA is duplex but only one strand is usually incorporated into RISC. [25, 26] RISC in association with miR binds to 3’ untranslated region (UTR) of target gene mRNA to inhibit protein translation or promote RNA degradation. Role of miRNARead MoreDr. Alwine : A Scientist Of Cancer Biology And An Investigator1048 Words   |  5 Pagesis a straight forward method. Several of the times this technique is used as a confirmation or verification of their research process. The Northern Blotting technique is a multipurpose procedure by allowing the usage of various types of probes. These probes include: radiolabeled and non-radiolabeled, oligonucleotides, such as primers, and in vitro transcribed RNA. This technique can also produce sequences then can be partial homology, unlike real time PCR or other procedures which can be used as hybridizationRead MoreUses of Nanotechnology1790 Words   |  8 PagesINTRODUCTION: In recent times, infectious diseases continue to pose a major healthcare issue in developing countries making it imperative to develop logical solutions for robust and rapid diagnosis and treatment of these infections. Traditional techniques suffer from limitations, including laborious specimen processing, bulky instrumentation, and slow result readout. In view of the urgency for sensitive, specific, robust and rapid diagnostics, numerous advancements have been made in the areaRead MoreCell Free DNA Case Study1819 Words   |  8 Pagesare various methods and technologies used for quantitative and qualitative analysis of ctDNAs, commonly used platforms to name a few are digital PCR (dPCR)283, droplet digital PCR (ddPCR)284, BEAMing285, 286, cancer personalized profiling by deep sequencing (CAPP-Seq)287, tagged amplicon deep sequencing (TAM-Seq)270, safe-sequencing (Safe-Seq)288, duplex sequencing289, integrated digital error suppression (iDES)-enhanced CAPP-Seq290, whole-genome sequencing (WGS)291, 292 and next-generation sequencing