Simultaneous identification of three highly pathogenic Eimeria species in rabbits using a multiplex PCR diagnostic assay based on ITS1-5.8S rRNA-ITS2 fragments

Yan, Wenchao; Wang, Wenlong; Wang, Tianqi; Suo, Xun; Qian, Weifeng; Wang, Shuai; Fan, Daiming

doi:10.1016/j.vetpar.2012.11.013

Cited by 29 publications

(20 citation statements)

References 15 publications

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“…A PCR has been developed (Oliveira et al, 2011) that differentiates between 11 of the different Eimeria species that infect the domestic rabbit. Smaller scale PCR for detection and differentiation between the more pathogenic species (E. intestinalis, E. flavicenens, and E. stiedae) has also been developed (Yan et al, 2013). Smaller scale PCR for detection and differentiation between the more pathogenic species (E. intestinalis, E. flavicenens, and E. stiedae) has also been developed (Yan et al, 2013).…”

Section: Intestinal Coccidiosismentioning

confidence: 99%

Biology and Diseases of Rabbits

Nowland

Brammer

García

et al. 2015

Laboratory Animal Medicine

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Section: Intestinal Coccidiosismentioning

confidence: 99%

Biology and Diseases of Rabbits

Nowland

Brammer

García

et al. 2015

Laboratory Animal Medicine

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“…Multiplex PCR assay enables the identification and detection of mixed infection in a single reaction that contains multiple primer pairs; it is a convenient, rapid, cost-effective, and high-efficiency method. So far, several multiplex PCR assays have been documented for the identification and detection of various parasite species [ 18 – 20 ]. Prior to the present study, there have been no reports of multiplex PCR assay for simultaneous detection of A. ceylanicum, A. caninum, and G. lamblia assemblage A.…”

Section: Introductionmentioning

confidence: 99%

A Multiplex PCR for Simultaneous Detection of Three Zoonotic ParasitesAncylostoma ceylanicum,A. caninum,andGiardia lambliaAssemblage A

et al. 2015

BioMed Research International

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Ancylostoma ceylanicum, A. caninum, and Giardia lamblia assemblage A are common intestinal parasites of dogs and cats; they can also infect humans, causing parasitic zoonoses. In this study, a multiplex PCR method was developed for simultaneous identification and detection of those three zoonotic parasites. Three pairs of specific primers were designed based on ITS sequence of A. ceylanicum and A. caninum and TPI gene of G. lamblia available in the GenBank. The multiplex PCR reaction system was established by optimizing the reaction condition, and a series of tests on the sensitivity, specificity, and clinical application were also conducted. Results showed that three target fragments were amplified specifically; the detection limit was 10 eggs for both A. ceylanicum and A. caninum, 72 pg DNA for G. lamblia. Of 112 clinical fecal samples, 34.8% and 17.8% samples were positive for A. caninum and A. ceylanicum, respectively, while only 2.7% samples were positive for G. lamblia assemblage A. It is concluded that the established multiplex PCR assay is a convenient, rapid, cost-effective, and high-efficiency method for molecular detection and epidemiological investigation of three zoonotic parasites.

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“…In addition to your target sequences, you need to download more sequences of related species to your target pathogens to align and fi nd accurately the polymorphic sites of your target sequences [ 6 ].…”

Section: Notesmentioning

confidence: 99%

“…The specifi c primer pairs for four pathogenic rabbit coccidia were designed using Primer-BLAST, and then analyzed mispriming and secondary structure potential with Oligo 6 and MFold bioinformatic software to develop a rapid, specifi c, and sensitive multiplex PCR differential assay [ 6 ].…”

Section: Introductionmentioning

confidence: 99%

Multiplex PCR Primer Design for Simultaneous Detection of Multiple Pathogens

Yan

2015

Methods in Molecular Biology

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Multiplex PCR provides a powerful tool for simultaneous detection and discrimination of multiple pathogens or different subtypes of a causative agent from humans, animals, and plants in a single reaction, and saves time and cost in the clinical diagnostic laboratory. Here, we describe the specific protocol of multiplex PCR primer design for simultaneous identification of more than one target from a same specimen. Different sizes of amplicons and similar T m values of primer sets are essential to successfully develop a feasible multiplex PCR assay.

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Simultaneous identification of three highly pathogenic Eimeria species in rabbits using a multiplex PCR diagnostic assay based on ITS1-5.8S rRNA-ITS2 fragments

Cited by 29 publications

References 15 publications

Biology and Diseases of Rabbits

Biology and Diseases of Rabbits

A Multiplex PCR for Simultaneous Detection of Three Zoonotic ParasitesAncylostoma ceylanicum,A. caninum,andGiardia lambliaAssemblage A

Multiplex PCR Primer Design for Simultaneous Detection of Multiple Pathogens

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