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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: A protocol which enables mitochondrial sequence information to be generated rapidly and automatically is described, likely to be of importance in forensic analysis where the DNA is too degraded or of insufficient quantity to be analysed by other techniques.
Abstract: Part of the human mitochondrial D-loop region was amplified by two successive rounds of polymerase chain reaction (PCR) amplification. In the second PCR reaction, nested primers were used, of which one contained the M13-21 universal primer sequence. By using nonequal concentrations of primers in the second amplification, single-stranded DNA was generated. This was then sequenced directly by the diodeoxy chain termination method using dye-labelled universal sequencing primers in conjunction with a fluorescence-based DNA sequencer. This enabled a 403-base-pair hypervariable segment of the D-loop region to be readily sequenced in a single reaction. This paper describes a protocol which enables mitochondrial sequence information to be generated rapidly and automatically. It is likely to be of importance in forensic analysis where the DNA is too degraded or of insufficient quantity to be analysed by other techniques.

62 citations

Journal ArticleDOI
TL;DR: The detection rate of a multiplex real-time polymerase chain reaction (PCR) test, with emphasis on epidemiology and seasonal distribution of the most common respiratory tract infections, supported the use of similar tests in routine clinical care all year round.
Abstract: Background: Nucleic acid amplification tests are increasingly being used to diagnose viral and bacterial respiratory tract infections. The high sensitivity of these tests affects our understanding of the epidemiology of respiratory tract infections. We have assessed the detection rate of a multiplex real-time polymerase chain reaction (PCR) test, with emphasis on epidemiology and seasonal distribution of the most common respiratory tract infections. Methods: Seven thousand eight hundred and fifty-three nasopharyngeal samples from 7220 patients (age range 0–98 y, median 22 y) obtained during 36 consecutive months (November 2006–October 2009), were analyzed with a multiplex PCR panel including influenza A (IfA) and B (IfB) virus, parainfluenza virus (PIV) 1–3, respiratory syncytial virus (RSV), human rhinovirus (HRV), human coronavirus (CoV) OC43, NL63, and 229E, human metapneumovirus (HMPV), adenovirus (AdV), enterovirus (EV), and 2 bacteria – Mycoplasma pneumoniae and Chlamydophila pneumoniae. Res...

62 citations

Journal ArticleDOI
TL;DR: A multiplex-PCR assay which uses genotype-specific primer pairs for HBV genotypes A-F is developed, which specifically amplified HBV DNA of the respective genotype, either in single or in multiplex -PCR.

62 citations

Journal ArticleDOI
TL;DR: The results of the collaborative exercise on typing of autosomal single nucleotide polymorphisms (SNP) for forensic genetic investigations in crime cases demonstrate that SNP typing with SBE, capillary electrophoresis and multicolour detection methods can be developed for forensic genetics.
Abstract: We report the results of an inter-laboratory exercise on typing of autosomal single nucleotide polymorphisms (SNP) for forensic genetic investigations in crime cases. The European DNA Profiling Group (EDNAP), a working group under the International Society for Forensic Genetics (ISFG), organised the exercise. A total of 11 European and one US forensic genetic laboratories tested a subset of a 52 SNP-multiplex PCR kit developed by the SNPforID consortium. The 52 SNP-multiplex kit amplifies 52 DNA fragments with 52 autosomal SNP loci in one multiplex PCR. The 52 SNPs are detected in two separate single base extension (SBE) multiplex reactions with 29 and 23 SNPs, respectively, using SNaPshot kit, capillary electrophoresis and multicolour fluorescence detection. For practical reasons, only the 29 SBE multiplex reaction was carried out by the participating laboratories. A total of 11 bloodstains on FTA cards including a sample of poor quality and a negative control were sent to the laboratories together with the essential reagents for the initial multiplex PCR and the multiplex SBE reaction. The total SNP locus dropout rate was 2.8% and more than 50% of the dropouts were observed with the poor quality sample. The overall rate of discrepant SNP allele assignments was 2.0%. Two laboratories reported 60% of all the discrepancies. Two laboratories reported all 29 SNP alleles in all 10 positive samples correctly. The results of the collaborative exercise were surprisingly good and demonstrate that SNP typing with SBE, capillary electrophoresis and multicolour detection methods can be developed for forensic genetics.

62 citations

Journal ArticleDOI
TL;DR: The diagnostic value of dual priming oligonucleotide (DPO)–based multiplex polymerase chain reaction (PCR) for the detection of BRAFV600E mutations in ultrasound‐guided fine‐needle aspiration biopsy (US‐FNAB) of thyroid nodules is evaluated.
Abstract: Background. To evaluate the diagnostic value of dual priming oligonucleotide (DPO)-based multiplex polymer- ase chain reaction (PCR) for the detection of BRAF V600E muta- tions in ultrasound-guided fine-needle aspiration biopsy (US- FNAB) of thyroid nodules. Methods. Our institutional review board approved this ret- rospective study, and informed consent was not required from patients. The 130 patients underwent US-FNAB to evaluate BRAF status in thyroid nodules. In FNAB washouts, DPO- based multiplex PCR, direct DNA sequencing, and PCR- restriction fragment length polymorphism (RFLP) were used to detect BRAF V600E. The diagnostic performance of these meth- ods was calculated. We compared cytologic results by BRAF status.

62 citations


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