scholarly journals Enhancing multiplex PCR efficiency using Hot Start dNTPs

BioTechniques ◽  
2009 ◽  
Vol 47 (5) ◽  
pp. 972-973 ◽  
Author(s):  
Tony Le ◽  
Elena Hidalgo Ashrafi ◽  
Natasha Paul
Keyword(s):  
Multiplex Pcr ◽  
Pcr Efficiency ◽  
Hot Start

scholarly journals Multiplex PCR: Optimization and Application in Diagnostic Virology

2000 ◽  
Vol 13 (4) ◽  
pp. 559-570 ◽  
Author(s):  
Elfath M. Elnifro ◽  
Ahmed M. Ashshi ◽  
Robert J. Cooper ◽  
Paul E. Klapper
Keyword(s):  
Multiplex Pcr ◽  
Disease Diagnosis ◽  
Test Design ◽  
Target Sequence ◽  
Infectious Agents ◽  
Simple Test ◽  
Multiplex Assays ◽  
Hot Start ◽  
Diagnostic Capacity ◽  
Diagnostic Virology

SUMMARY PCR has revolutionized the field of infectious disease diagnosis. To overcome the inherent disadvantage of cost and to improve the diagnostic capacity of the test, multiplex PCR, a variant of the test in which more than one target sequence is amplified using more than one pair of primers, has been developed. Multiplex PCRs to detect viral, bacterial, and/or other infectious agents in one reaction tube have been described. Early studies highlighted the obstacles that can jeopardize the production of sensitive and specific multiplex assays, but more recent studies have provided systematic protocols and technical improvements for simple test design. The most useful of these are the empirical choice of oligonucleotide primers and the use of hot start-based PCR methodology. These advances along with others to enhance sensitivity and specificity and to facilitate automation have resulted in the appearance of numerous publications regarding the application of multiplex PCR in the diagnosis of infectious agents, especially those which target viral nucleic acids. This article reviews the principles, optimization, and application of multiplex PCR for the detection of viruses of clinical and epidemiological importance.

Improvement of Multiplex PCR Efficiency by Using The MPprimer to Design Primers and Optimizing The Amplification Conditions*

2010 ◽  
Vol 37 (3) ◽  
pp. 342-346 ◽  
Author(s):  
Wen WANG ◽  
Wu-Bin QU ◽  
Zhi-Yong SHEN ◽  
Chang-Hong REN ◽  
Hu-Qi LIU ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Pcr Efficiency

A multiplex PCR-SSCP test to genotype bovine β-casein alleles A1 , A2 , A3 , B, and C

1999 ◽  
Vol 30 (4) ◽  
pp. 322-323 ◽  
Author(s):  
A. Barroso ◽  
S. Dunner ◽  
J. Ca·ón
Keyword(s):  
Multiplex Pcr

Multiplex PCR zum Nachweis disseminierter Tumorzellen (DTC) im Blut von Patientinnen mit Mammakarzinom

2005 ◽  
Vol 127 (03) ◽  
Author(s):  
J Rom ◽  
A Schneeeweiss ◽  
V Zieglschmid ◽  
C Hollmann ◽  
O Böcher ◽  
...  
Keyword(s):  
Multiplex Pcr

Usefulness of a Multiplex PCR for Detection of Diarrheagenic Escherichia coli in a Diagnostic Microbiology Laboratory Setting

2016 ◽  
Vol 1 (2) ◽  
pp. 38-42 ◽  
Author(s):  
Khairun Nessa ◽  
Dilruba Ahmed ◽  
Johirul Islam ◽  
FM Lutful Kabir ◽  
M Anowar Hossain
Keyword(s):  
Escherichia Coli ◽  
Multiplex Pcr ◽  
Clinical Laboratory ◽  
Proteinase K ◽  
Microbiology Laboratory ◽  
Pcr Assay ◽  
E Coli ◽  
Diarrheagenic Escherichia Coli ◽  
Multiplex Pcr Assay ◽  
Diagnostic Microbiology

A multiplex PCR assay was evaluated for diagnosis of diarrheagenic Escherichia coli in stool samples of patients with diarrhoea submitted to a diagnostic microbiology laboratory. Two procedures of DNA template preparationproteinase K buffer method and the boiling method were evaluated to examine isolates of E. coli from 150 selected diarrhoeal cases. By proteinase K buffer method, 119 strains (79.3%) of E. coli were characterized to various categories by their genes that included 55.5% enteroaggregative E. coli (EAEC), 18.5% enterotoxigenic E. coli (ETEC), 1.7% enteropathogenic E. coli (EPEC), and 0.8% Shiga toxin-producing E. coli (STEC). Although boiling method was less time consuming (<24 hrs) and less costly (<8.0 US $/ per test) but was less efficient in typing E. coli compared to proteinase K method (41.3% vs. 79.3% ; p<0.001). The sensitivity and specificity of boiling method compared to proteinase K method was 48.7% and 87.1% while the positive and negative predictive value was 93.5% and 30.7%, respectively. The majority of pathogenic E. coli were detected in children (78.0%) under five years age with 53.3% under one year, and 68.7% of the children were male. Children under 5 years age were frequently infected with EAEC (71.6%) compared to ETEC (24.3%), EPEC (2.7%) and STEC (1.4%). The multiplex PCR assay could be effectively used as a rapid diagnostic tool for characterization of diarrheagenic E. coli using a single reaction tube in the clinical laboratory setting.Bangladesh J Med Microbiol 2007; 01 (02): 38-42

EVALUATION OF THE ISOLATION AND DETECTION METHODS FOR SALMONELLA SPP. FROM EGG SHELL CONTAMINATION USING MULTIPLEX PCR.

2015 ◽  
Vol 14 (1) ◽  
pp. 282-288
Author(s):  
Israa Adnan Ibraheam Al-Baghdady ◽  
Ashwak Bassim Jassim ◽  
Zainab Khudher Ahmed
Keyword(s):  
Multiplex Pcr ◽  
Detection Methods ◽  
Salmonella Spp ◽  
Egg Shell
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