10 results about "Multiplex polymerase chain reaction" patented technology

Method for evaluating a Multiplex Polymerase Chain Reaction (Multiplex PCR) primer and probe set, comprising the steps of: a) generating all potential primers and probes from a pool of candidate sequences; b) initiating a global optimization algorithm to generate primer and probe sets from the generated pool; c) calculating an Intramolecular Score (SI), a Free Primer / Probe Concentration Score (SF), a Dimerization Score (SD), and a Hybridization Efficiency Score (SH) for each primer and probe of the set; d) calculating a Final Oligo Score (SO) for each primer and probe of the set b); e) calculating a Total Set Score (ST) as the sum of all Final Oligo Scores (SO) of each primer and probe of the set; and f) ranking each primer and probe set according to its Total Set Score (ST). There is also describe a computer program product configured for performing said method.

The application belongs to the technical field of gene detection and molecular diagnosis, and particularly relates to a kit for detecting a pathogenic gene of phenylketonuria and application thereof. The kit comprises a primer pair for amplifying specific exons and splicing regions of a phenylalanine hydroxylase gene, and specific probes for detecting hot spot mutations and deletion / repetition variations of the gene. The application can detect genetic variations related to phenylketonuria in one time, quickly and accurately by combining optimized multiplex polymerase chain reaction with high-throughput sequencing or gene chip technology, and covers various known hot spot mutations and copy number variations including c.1222C>T, c.1068-11G>A, c.728G>A and c.1162G>A. The kit has high detection sensitivity and strong specificity, and is suitable for positive recall diagnosis of neonatal phenylketonuria screening, genetic diagnosis of suspected patients, carrier screening and prenatal diagnosis, and provides an efficient tool for precise prevention and control of phenylketonuria.

Described herein are compositions, methods, and kits for detecting wound pathogens. One embodiment described herein is primer pairs and probes for individual, simultaneous, or sequential multiplex polymerase chain reaction (PCR) based assays for the detection of Acinetobacter baumannii, Bacteroides fragilis, Clostridium perfringens, Enterobacter cloacae, Enterococcus faecalis, Enterococcus faecium, Escherichia coli, Klebsiella pneumoniae, Klebsiella Oxytoca, Klebsiella aerogenes, Proteus mirabilis, Proteus vulgaris, Pseudomonas aeruginosa, Serratia marcescens, Staphylococcus aureus, Streptococcus agalactiae, Streptococcus dysgalactiae, Streptococcus pyogenes, Streptococcus anginosus, Staphylococcus lugdunensis, Providencia stuartii, Stenotrophomonas maltophilia, Morganella morganii, Citrobacter koseri, Citrobacter freundii, Citrobacter braakii, Peptoniphilus asaccharolyticus, Peptoniphilus harei, Peptoniphilus ivorii, Peptostreptococcus anaerobius, Finegoldia magna, Clostridium septicum, Paeniclostridium sordellii (Clostridium sordellii), Clostridium novyi A, B, Clostridium histolyticum, Staphylococcus epidermidis, Staphylococcus haemolyticus, Corynebacterium striatum, Candida albicans, Candida auris, Candida glabrata, Candida krusei, Candida parapsilosis, or Candida tropicalis.

The present disclosure provides compositions, methods, and kits for detecting guaiacol-producing microorganisms, in particular, Thermophilic Acidophilic Bacteria (TAB) in foodstuff. In one example, a method comprises: (a) contacting a set of oligonucleotides with the sample, wherein the set of oligonucleotides comprising: a first collection of primers targeted for nucleic acids of a guaiacol-producing microorganism; and a second collection of primers targeted for nucleic acids of Alicyclobacillus acidoterrestris; (b) amplifying DNA in the sample with the said set of oligonucleotides under a multiplex polymerase chain reaction (PCR); and (c) determining the presence of PCR products of step (b), wherein the presence of a PCR product in the sample is indicative of contamination of the sample by both guaiacol-producing microorganism and Alicyclobacillus acidoterrestris.

This invention discloses a strategy for rapid detection of isoniazid resistance in Mycobacterium tuberculosis based on multiplex polymerase chain reaction (Multiplex PCR) using local rTth DNA polymerase and interpretation of amplification patterns. This strategy utilizes a specific primer combination that allows simultaneous amplification of the control fragment and the target fragment of the katG gene codon 315 mutation in a single PCR reaction. The amplification products are analyzed by 1% (w / v) agarose gel electrophoresis and interpreted based on the resulting DNA banding pattern to distinguish sensitive isolates, isolates carrying the katG315 mutation causing isoniazid resistance, and invalid test results. The use of local rTth DNA polymerase provides an alternative thermostable enzyme to support national diagnostic raw material independence without changing the detection principle.This strategy allows for rapid identification of katG315 mutations associated with isoniazid resistance without the need for DNA sequencing or further molecular analysis. The invention provides a simple, rapid, specific, easily interpretable, and cost-effective molecular diagnostic method to support the early detection of isoniazid-resistant tuberculosis.

The present invention provides a novel means for detecting various variants of carbapenemase-producing Enterobacterales (CPE) in which carbapenemase is an enzyme breaking down carbapenem antibiotics. The primer sets of the present invention are capable of detecting, with high accuracy in silico, CPE-associated genes such as KPC, NDM, OXA-48-family, GES, VIM-2-like, IMP G1, and variants thereof. Even when the primer sets are combined for multiplex polymerase chain reaction (PCR), nonspecific bands do not appear at the same annealing temperature, with the generation of PCR amplification products of different sizes, allowing detection of various CPE-related genes and variants thereof in a single PCR reaction. Furthermore, the use of probes that specifically bind to each PCR amplification product enables application in real-time PCR, thus providing a highly accurate and specific means for detecting carbapenemase producing Enterobacterales (CPE) strains.

Described herein are compositions, methods, and kits for detecting antibiotic resistance genes in a sample, such as a wound swab. One embodiment described herein is primer pairs and probes for individual or multiplex polymerase chain reaction (PCR) based assays for the detection of one or more antibiotic resistance gene targets comprising resistance to molecularly characterized extended-spectrum β-lactamases (MESBLs); extended-spectrum β-lactamases (ESBLs); carbapenemase; AmpC β-lactamase; β-lactamase; lincosamide, macrolide, streptogramin; trimethoprim; macrolides; methicillin; colistin; sulfonamide; tetracycline; vancomycin; nitroimidazole; quinolone; or aminoglycoside.

The invention relates to the fields of biotechnology, molecular biology and clinical diagnosis, in particular to a method for efficiently enriching aerobic bacteria and anaerobic bacteria by utilizing a mixture of multiple microorganism enrichment magnetic bead reagents and combining with a multiplex polymerase chain reaction (PCR) technology so as to improve the detection efficiency of a low-concentration microorganism sample. The method is suitable for pathogenic microorganism detection in medical examination.

Described herein are compositions, methods, and kits for detecting diarrhea-causing pathogens from a patient, food, or environmental sample. One embodiment described herein is a primer pair and probe for a multiplex polymerase chain reaction (PCR)-based assay for the detection of diarrhea-causing pathogens, such as Campylobacter spp., Salmonella spp., Shigella spp. / Enteroinvasive Escherichia coli (EIEC), Escherichia coli stx1 / Shiga toxin A, or Escherichia coli stx2 / Shiga toxin B. Other embodiments include methods and kits for detecting diarrhea-causing pathogens.

This document describes compositions, methods, and kits for detecting diarrheal pathogens from patient, food, or environmental samples. One embodiment described herein is primer pairs and probes for a multiplex polymerase chain reaction (PCR)-based assay for detecting gastrointestinal pathogens such as adenovirus, astrovirus, rotavirus, norovirus, zaruzin virus, Campylobacter species, Clostridium difficile, enteroaggregative Escherichia coli (EAEC), enteropathogenic Escherichia coli (EPEC), enterotoxin-producing Escherichia coli (ETEC), Shigella-like toxin-producing Escherichia coli (STEC), Shigella spp. / enteroinvasive Escherichia coli (EIEC), Escherichia coli O157, Listeria monocytogenes, Shigella-like bacteria, Salmonella spp., Yersinia enterocolitica, Vibrio species, Cryptosporidium species, Cyclospora kayanoides, Entamoeba histolytica, Giardia lamblia, or Giardia lamblia. Other embodiments include methods and kits for detecting gastrointestinal pathogens.