A number of review and research articles have provided detailed descriptions of the key parameters that may influence the performance of standard (uniplex) PCR (17, 57, 88, 91, 112). The accuracy of this approach was confirmed by examining cervical scrapings from 44 patients. Uniplex PCR would be an impractical approach for rapid screening of hundreds of specimens per day for a range of transfusion-transmitted viruses. Development and application of a multiplex PCR assay for detection of the Clostridium perfringens enterotoxin-encoding genes cpe and becAB J Microbiol Methods. In the second round of amplification, five additional subtype-specific primers and two universal non-group-specific primers are added to generate two to four DNA fragments of defined sizes indicative for the subtype. Enter multiple addresses on separate lines or separate them with commas. In addition, two uniplex PCRs were used for detection of CMV and JC virus. The method proved to be a simple, economical, and reliable tool for detection of HPV infection. Conducting polymers on microelectronic devices as tools for biological analyses. The latter may be achieved by using primer pairs that result in PCR products that can be separated and clearly visualized using gel electrophoresis or hybridization probes with maximum specificity. In modern-day science, the present method is widely used in microbial detection and identification, and virology studies. Direct detection of Sabin poliovirus vaccine strains in stool specimens of first-dose vaccinees by a sensitive reverse transcription-PCR method. More recently, a simple multiplex PCR utilizing three primer pairs for amplification of HPV-16, -18, and -33 in combination with a colorimetric microplate hybridization as a post-PCR detection system has been developed (35). Evaluation of celiac disease biopsies for adenovirus 12 and using a multiplex polymerase chain reaction. Comparison of primers and optimization of PCR conditions for detection of. Multiplex polymerase chain reaction (Multiplex PCR) is a widespread molecular biology technique for amplification of multiple targets in a single PCR experiment. In a total of 195 stool samples collected from 26 vaccinees following administration of the first dose of the trivalent oral vaccine, the multiplex PCR was more sensitive than culture for the detection of poliovirus types 1, 2, and 3. 2017 Jan;116(1):217-224. doi: 10.1007/s00436-016-5282-0. PCR conditions, such as compatibility among the primers within the reaction mixture such that there is no interference, is of great technical importance. The test was further modified to include a reverse transcription step and primer pairs to detect enterovirus cDNA (12). In patients for whom a diagnosis was confirmed at autopsy, the test was positive for HSV-1 or -2 for 19 patients (2%), producing a sensitivity and specificity of 100 and 99.6%, respectively. Mutation analysis of DMD. This may be achieved through the utilization of primers with nearly identical optimum annealing temperatures (primer length of 18 to 30 bp or more and a GC content of 35 to 60% may prove satisfactory) and should not display significant homology either internally or to one another (17, 26, 43). Mutation studies of the dystrophin gene are focused on detecting deletions or duplications of one or more exons, and multiplex PCR that amplifies selected deletion-prone exons has been used as the most efficient method of mutation detection [34,35].Mutations in dystrophin protein disrupt the open reading frame and prevent the full translation … Shahdoust S, Niyyati M, Haghighi A, Azargashb E, Khataminejad MR. Gastroenterol Hepatol Bed Bench. The PCR Markets 7.1 PCR - Global Market Overview by Country. Enterovirus was detected in 49 patients, HSV was detected in 3 patients, VZV was detected in 6 patients, CMV was detected in 12 patients, EBV was detected in 2 patients, CMV and HSV were detected in one AIDS patient with encephalitis, and CMV and EBV were detected in another AIDS patient with polyradiculomyelitis. Epidemiol Bulletin. Epub 2016 Jun 10. In a multiplexing assay, more than one target sequence is amplified by using multiple primer pairs in the same reaction mixture. The same multiplex PCR was used to evaluate swab specimens from 38 sequential patients with genital ulcer disease who received clinical diagnoses and syndromic treatment (7). Laboratory diagnostic techniques for Entamoeba species. In heart transplant recipients a hot start nested multiplex PCR was used to evaluate the possible reactivation of HHV-6 and HHV-7 (67). Multiplex, real-time RT-PCR is a similar method, allowing simultaneous quantification of several RNA targets in the same reaction. For example, in the multiplex PCR for the dystrophin gene (nine genomic targets), a TaqDNA polymerase concentration (with an appropriate increase in MgCl2 concentration) four to five times greater than that required in uniplex PCR was necessary to achieve optimal nucleic acid amplification (19). An introduction to PCR primer design and optimisation of amplification reactions. Automation of molecular genetic methods. Transcription of the human cytomegalovirus natural killer decoy gene, UL18. Polymerase chain reaction in transfusion medicine. All samples from patients with condyloma acuminata contained HPV DNA but mainly contained HPV-6/11 (87.5%), while high-risk HPV had a low prevalence rate (6.5%). The test proved to have a high diagnostic utility for detection of CMV variants with maximal sensitivity and specificity. Variation in concentrations of reaction components above those used in uniplex PCR probably reflects the competitive nature of the PCR process. 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