Topic
Multiplex polymerase chain reaction
About: Multiplex polymerase chain reaction is a research topic. Over the lifetime, 6409 publications have been published within this topic receiving 221244 citations.
Papers published on a yearly basis
Papers
More filters
TL;DR: A nested multiplex PCR was developed for genotyping of bovine viral diarrhea viruses (BVDVs) that could detect as little as 3 50% tissue culture infective doses of BVDV per ml and typed 42 out of 42 cell culture isolates.
Abstract: A nested multiplex PCR was developed for genotyping of bovine viral diarrhea viruses (BVDVs). The assay could detect as little as 3 50% tissue culture infective doses of BVDV per ml and typed 42 out of 42 cell culture isolates. BVDV was also successfully typed, with or without RNA extraction, from all 27 whole-blood samples examined from 22 carriers or probable carriers and 5 experimentally infected cattle.
72 citations
TL;DR: A novel 10-primer pair multiplex PCR and reverse line blot (mPCR/RLB) hybridization assay was developed for simultaneous detection and serotype identification of all nine GBS serotypes and was more convenient and practical than either conventional serotyping or individual serotype-specific PCR.
Abstract: Streptococcus agalactiae (group B streptococcus, GBS) is an important cause of sepsis in neonates and their mothers, and the elderly and immunocompromised patients. Ongoing surveillance to monitor GBS serotype distribution is needed to guide the development and assess the feasibility of GBS conjugate vaccines. The authors previously developed a molecular serotype identification method based on serotype-specific PCR and partial sequencing of cps genes. In this study, a novel 10-primer pair multiplex PCR and reverse line blot (mPCR/RLB) hybridization assay was developed for simultaneous detection and serotype identification of all nine GBS serotypes. For all 316 GBS isolates tested the mPCR/RLB results corresponded with those of conventional serotyping and individual serotype-specific PCR, and the method was more convenient and practical than either alternative.
71 citations
TL;DR: Results obtained suggest this technique could be applied as a sole diagnostic tool in several clinical settings in which herpesviral infection is suspected and differential diagnosis required, avoiding the need to test specimens by separate PCR methods.
71 citations
TL;DR: How PCR has had an impact on molecular biology, human genetics, infectious and genetic disease diagnosis, forensic science, and evolutionary biology is discussed.
Abstract: This paper discusses the polymerase chain reaction (PCR) an in-vitro method of amplifying DNA sequences. Beginning with DNA of any origin- bacterial, viral, plant, or animal- PCR can increase the amount of a DNA sequence hundreds of millions to billions of times. The procedure can amplify a targeted sequence even when it makes up less than one part in a million of the total initial sample. PCR is an enzymatic process that is carried out in discrete cycles of amplification, each of which can double the amount of target DNA in the sample. Thus, n cycles can produce 2{sup n} times as much target as was present to begin with. This paper discusses how PCR has had an impact on molecular biology, human genetics, infectious and genetic disease diagnosis, forensic science, and evolutionary biology.
71 citations
TL;DR: It is concluded that the straightforward procedure substantially expands the applicability of the massive parallel sequencers for sequencing projects of a moderate number of amplicons with typical applications in resequencing exons in positional or functional candidate regions and molecular genetic diagnostics.
Abstract: We evaluated multiplex PCR amplification as a front-end for high-throughput sequencing, to widen the applicability of massive parallel sequencers for the detailed analysis of complex genomes. Using multiplex PCR reactions, we sequenced the complete coding regions of seven genes implicated in peripheral neuropathies in 40 individuals on a GS-FLX genome sequencer (Roche). The resulting dataset showed highly specific and uniform amplification. Comparison of the GS-FLX sequencing data with the dataset generated by Sanger sequencing confirmed the detection of all variants present and proved the sensitivity of the method for mutation detection. In addition, we showed that we could exploit the multiplexed PCR amplicons to determine individual copy number variation (CNV), increasing the spectrum of detected variations to both genetic and genomic variants. We conclude that our straightforward procedure substantially expands the applicability of the massive parallel sequencers for sequencing projects of a moderate number of amplicons (50-500) with typical applications in resequencing exons in positional or functional candidate regions and molecular genetic diagnostics.
71 citations