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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.


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Journal ArticleDOI
TL;DR: The fast multiplex PCR approach has the potential to reduce process time and cost for STR analysis and enables development of a fully integrated microfluidic forensic DNA analysis system.
Abstract: The time required for short tandem repeat (STR) amplification is determined by the temperature ramp rates of the thermal cycler,the components of the reaction mix, and the properties of the reaction vessel. Multiplex amplifications in microfluidic biochip-based and conventionaltube-based thermal cyclers have been demonstrated in 17.3 and 19 min, respectively. Optimized 28-cycle amplification protocols generated alleleswith signal strengths above calling thresholds, heterozygous peak height ratios of greater than 0.65, and incomplete nontemplate nucleotide additionand stutter of less than 15%. Full CODIS-compatible profiles were generated using the Profiler Plus ID, COfiler and Identifiler primer sets. PCR per-formance over a wide range of DNA template levels from 0.006 to 4 ng was characterized by separation and detection on a microfluidic electropho-resis system, Genebench-FXTM. The fast multiplex PCR approach has the potential to reduce process time and cost for STR analysis and enablesdevelopment of a fully integrated microfluidic forensic DNA analysis system

66 citations

Journal ArticleDOI
TL;DR: A qualitative PCR-based multiplex method for simultaneous detection and identification of four genetically modified maize lines: Bt11, MON810, T25, and GA21, based on the use of five primers directed to specific sequences in these insertion events.
Abstract: The number of cultured hectares and commercialized genetically modified organisms (GMOs) has increased exponentially in the past 9 years Governments in many countries have established a policy of labeling all food and feed containing or produced by GMOs Consequently, versatile, laboratory-transferable GMO detection methods are in increasing demand Here, we describe a qualitative PCR-based multiplex method for simultaneous detection and identification of four genetically modified maize lines: Bt11, MON810, T25, and GA21 The described system is based on the use of five primers directed to specific sequences in these insertion events Primers were used in a single optimized multiplex PCR reaction, and sequences of the amplified fragments are reported The assay allows amplification of the MON810 event from the 35S promoter to the hsp intron yielding a 468 bp amplicon Amplification of the Bt11 and T25 events from the 35S promoter to the PAT gene yielded two different amplicons of 280 and 177 bp, respectively, whereas amplification of the 5' flanking region of the GA21 gave rise to an amplicon of 72 bp These fragments are clearly distinguishable in agarose gels and have been reproduced successfully in a different laboratory Hence, the proposed method comprises a rapid, simple, reliable, and sensitive (down to 005%) PCR-based assay, suitable for detection of these four GM maize lines in a single reaction

66 citations

Journal ArticleDOI
TL;DR: If the primary aim of PCR diagnostics is to decrease inappropriate empirical treatment and improve patient outcome, detection should focus on those pathogens or resistance determinants that are not covered by guideline-recommended treatment regimens and that have been identified as the major cause of inappropriate treatment according to current studies.
Abstract: Polymerase chain reaction (PCR)-based techniques allow more rapid and sensitive detection of pathogens compared with conventional blood culture. Nevertheless, the climate of opinion of relevant studies is that currently PCR can supplement but not replace blood culture. In numerous studies, combined detection rate of both methods was significantly higher compared with PCR or blood culture alone. Also, complete determination of antibiotic resistance can currently be performed only by blood culture. Further increase of the panel of multiplex PCR is complicated, because the vast majority of sepsis pathogens are already included, primer interactions leading to primer heteromers limit the amount of targets detectable within one PCR tube, and an array of too many individual PCR reactions for investigation of a single specimen leads to high cost and workload. Except for diagnostics of patients in whom unusual, not culturable, or fastidious pathogens are detected more often, such as immunosuppressed patients with suspected parasitic infection, etc., it might even not be necessary to further increase the spectrum of detectable species. If the primary aim of PCR diagnostics is to decrease inappropriate empirical treatment and improve patient outcome, detection should focus on those pathogens or resistance determinants that are not covered by guideline-recommended treatment regimens and that have been identified as the major cause of inappropriate treatment according to current studies. In our opinion, such a narrower assay is more cost effective, may achieve higher accuracy due to reduced intratest interference, and would better address current and emerging clinical needs.

66 citations

Journal ArticleDOI
TL;DR: The multiplex PCR amplification systems offer a non-isotopic method for rapid, simple and accurate analysis of STR loci, which has immediate and valuable application in forensic analysis, paternity determination, tissue culture strain identification and bone marrow transplantation studies.
Abstract: Multiplex PCR amplification systems were developed using well-characterized, polymorphic short tandem repeat (STR) loci. Eight loci utilized in the multiplex amplifications included HUMCSF1PO, HUMTPOX, HUMTH01, HUMVWFA31, HUMF13A01, HUMFESFPS, HUMBFXIII and HUMLIPOL. From this list, three or four non-overlapping loci were simultaneously amplified, separated by denaturing polyacrylamide gel electrophoresis and visualized using silver stain or fluorescence detection. The multiplex PCR amplification systems offer a non-isotopic method for rapid, simple and accurate analysis of STR loci. This high-throughput method for DNA identification has immediate and valuable application in forensic analysis, paternity determination, tissue culture strain identification and bone marrow transplantation studies.

66 citations

Journal ArticleDOI
TL;DR: Quantitative fluorescence multiplex polymerase chain reaction is a reliable and rapid method that allows prenatal diagnosis of the major numeric chromosomal abnormalities to be performed within 24 hours.

66 citations


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Performance
Metrics
No. of papers in the topic in previous years
YearPapers
2023275
2022448
2021172
2020176
2019221
2018220