A multiplex PCR assay for the simultaneous detection and discrimination of the sevenEimeriaspecies that infect domestic fowl

Parasitology ◽  
2003 ◽  
Vol 127 (4) ◽  
pp. 317-325 ◽  
Author(s):  
S. FERNANDEZ ◽  
A. H. PAGOTTO ◽  
M. M. FURTADO ◽  
Â. M. KATSUYAMA ◽  
A. M. B. N. MADEIRA ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Domestic Fowl ◽  
Detection Threshold ◽  
Simultaneous Detection ◽  
Cost Effective ◽  
Pcr Assay ◽  
Multiplex Pcr Assay ◽  
Sensitivity Tests ◽  
Different Sources ◽  
Single Tube Reaction

This study reports the development of a novel multiplex PCR assay based on SCAR (Sequence-Characterised Amplified Region) markers for the simultaneous diagnosis of the 7Eimeriaspecies that infect domestic fowl. Primer pairs specific for each species were designed in order to generate a ladder of amplification products ranging from 200 to 811 bp. Sensitivity tests for each species were carried out, showing a detection threshold of 1–5 pg, which corresponds approximately to 2–8 sporulated oocysts. Distinct isolates of the 7Eimeriaspecies from different geographical sources were tested and successfully detected by the assay. All the species were amplified homogeneously, whether or not one of them was present in a high quantity, indicating that there was no cross-interference. The assay was also tested with different sources ofTaqDNA polymerase and thermocycler models, confirming the high reproducibility of the reaction. The economy of consumables and labour represented by a single-tube reaction greatly facilitates the molecular diagnosis of a large number of samples, making it appropriate for field epizootiological surveys. We propose the use of this multiplex PCR assay as a rapid and cost-effective diagnostic method for the detection and discrimination of the 7Eimeriaspecies that infect domestic fowl.

A new one-step multiplex PCR assay for simultaneous detection and identification of avian haemosporidian parasites

2018 ◽  
Vol 118 (1) ◽  
pp. 191-201 ◽  
Author(s):  
Arif Ciloglu ◽  
Vincenzo A. Ellis ◽  
Rasa Bernotienė ◽  
Gediminas Valkiūnas ◽  
Staffan Bensch
Keyword(s):  
Multiplex Pcr ◽  
Simultaneous Detection ◽  
Pcr Assay ◽  
Haemosporidian Parasites ◽  
Multiplex Pcr Assay ◽  
Detection And Identification ◽  
One Step

Multiplex PCR Detection of Enterotoxin Genes in Aeromonas spp. from Suspect Food Samples in Northern Taiwan

2008 ◽  
Vol 71 (10) ◽  
pp. 2094-2099 ◽  
Author(s):  
YU-CHANG CHANG ◽  
JAN-YI WANG ◽  
AMMAIYAPPAN SELVAM ◽  
SHU-CHEN KAO ◽  
SHANG-SHYNG YANG ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Diarrheal Disease ◽  
Simultaneous Detection ◽  
Food Poisoning ◽  
Food Samples ◽  
Pcr Assay ◽  
Multiplex Pcr Assay ◽  
Enterotoxin Genes ◽  
Causative Agents ◽  
Northern Taiwan

Aeromonads possess an array of virulence factors and are causative agents of a number of human infections. Among them, genes of one cytotoxic (Act) and two cytotonic (Alt, Ast) enterotoxins are implicated in a human diarrheal disease. A rapid, specific, simultaneous detection of these enterotoxin genes in suspected food poisoning samples is not yet reported. Hence, a multiplex PCR assay was designed to amplify the cytotoxic (act), heat-labile cytotonic (alt), and heat-stable cytotonic (ast) enterotoxin genes of aeromonads. The PCR assay was tested with 133 Aeromonas spp. isolated from suspect food poisoning samples and retail samples of poultry and fish from wet markets in and around Taipei, Northern Taiwan. The Aeromonas spp. isolates were divided into six genotypes based on absence or presence of one or more enterotoxin genes. Of these 133 isolates, Aeromonas caviae (52.5%) and Aeromonas hydrophila (43.4%) were the most frequently isolated species from food poisoning samples and retail samples, respectively. Among the species, A. hydrophila had a significantly higher proportion for harboring three enterotoxin genes than had the others, whereas Aeromonas encheleia, considered a nonpathogen, was found harboring three enterotoxin genes. The multiplex PCR assays are rapid and specific, and provide a useful tool for the detection and genotyping of enterotoxin genes of aeromonads.

Development and application of multiplex PCR assay for the simultaneous detection of Babesia vogeli, Ehrlichia canis and Hepatozoon canis in dogs

Acta Tropica ◽  
2020 ◽  
Vol 212 ◽  
pp. 105713
Author(s):  
Navpreet Kaur ◽  
Harkirat Singh ◽  
Payal Sharma ◽  
Niraj Kumar Singh ◽  
Neeraj Kashyap ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Simultaneous Detection ◽  
Ehrlichia Canis ◽  
Hepatozoon Canis ◽  
Pcr Assay ◽  
Multiplex Pcr Assay ◽  
Babesia Vogeli

scholarly journals Species-Specific Identification of Human Adenoviruses by a Multiplex PCR Assay

2000 ◽  
Vol 38 (11) ◽  
pp. 4114-4120 ◽  
Author(s):  
WanHong Xu ◽  
Mike C. McDonough ◽  
Dean D. Erdman
Keyword(s):  
Multiplex Pcr ◽  
Human Adenovirus ◽  
Cost Effective ◽  
Pcr Assay ◽  
Multiplex Pcr Assay ◽  
Amplicon Size ◽  
Species Specific ◽  
Fiber Gene ◽  
Amplification Reaction

A multiplex PCR assay was developed by using primers to the fiber gene that could differentiate human adenovirus (Ad) species A through F in a single amplification reaction. The assay correctly identified the species of all 49 recognized Ad prototype strains as well as 180 geographically and temporally diverse Ad field isolates. Ad serotype 6 (Ad6) (species C), Ad16 (species B), Ad31 (species A), and Ad40 and Ad41 (species F) could also be distinguished by amplicon size within each respective species. In comparison, a previously described Ad species-specific multiplex PCR assay that used primers to the Ad hexon gene gave equivocal results with several serotypes of species B, whereas our multiplex assay amplified all species B serotypes equally well. Our multiplex PCR assay will permit rapid, accurate, and cost-effective classification of Ad isolates.

A gold nanoparticles-assisted multiplex PCR assay for simultaneous detection of Salmonella typhimurium, Listeria monocytogenes and Escherichia coli O157:H7

2020 ◽  
Vol 12 (2) ◽  
pp. 212-217 ◽  
Author(s):  
Juan Du ◽  
Shujing Wu ◽  
Liyuan Niu ◽  
Junguang Li ◽  
Dianbo Zhao ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Gold Nanoparticles ◽  
Listeria Monocytogenes ◽  
Salmonella Typhimurium ◽  
Multiplex Pcr ◽  
Simultaneous Detection ◽  
Escherichia Coli O157 ◽  
Pcr Assay ◽  
Multiplex Pcr Assay ◽  
Pcr Product

Unfunctionalized flower-shaped AuNPs is used as colorimetric sensor for PCR product detection by naked eyes.

A rapid and reliable multiplex PCR assay for simultaneous detection of fourteen animal species in two tubes

2019 ◽  
Vol 295 ◽  
pp. 395-402 ◽  
Author(s):  
Jinchun Li ◽  
Jiapeng Li ◽  
Suigen Xu ◽  
Suyue Xiong ◽  
Junna Yang ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Animal Species ◽  
Simultaneous Detection ◽  
Pcr Assay ◽  
Multiplex Pcr Assay

Development of multiplex PCR for rapid detection of metallo-β-lactamase genes in clinical isolates of Acinetobacter baumannii

2020 ◽  
Author(s):  
Reza Ranjbar ◽  
Shahin Zayeri ◽  
Amir Mirzaie
Keyword(s):  
Acinetobacter Baumannii ◽  
Multiplex Pcr ◽  
Rapid Detection ◽  
Simultaneous Detection ◽  
Cross Reactivity ◽  
Clinical Isolates ◽  
Clinical Samples ◽  
Pcr Assay ◽  
Multiplex Pcr Assay

Background and Objectives: Acinetobacter baumannii has been known as a major pathogen causing nosocomial infec- tions. The aim of this study was to develop multiplex PCR for rapid and simultaneous detection of metallo-β-lactamase (MBL) genes in clinical isolates of A. baumannii. Materials and Methods: In this study, we used three sets of primers to amplify the MBL genes including bla        ,     bla   and bla   OXA-48 . The multiplex PCR assay was optimized for rapid and simultaneous detection of MBL genes in A. bau-   OXA-23   NDM   mannii strains recovered from clinical samples. Results: A. baumannii strains recovered from clinical samples were subjected to the study. The multiplex PCR produced 3   OXA-48   OXA-23   bands of 501 bp for bla        , 744 bp for bla observed in multiplex PCR.   OXA-48   and 623 bp for bla   NDM   genes. In addition to, no any cross-reactivity was   Conclusion: Based on obtained data, the multiplex PCR had a good specificity without any cross reactivity and it appears that the multiplex PCR is reliable assay for simultaneous detection of MBL genes in A. baumannii strains.  

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