Development and evaluation of a quantitative PCR assay for detection of Hepatozoon sp.

被引：31

作者：

Criado-Fornelio, A. ^{[1
]}

Buling, A. ^{[1
]}

Cunha-Filho, N. A. ^{[2
]}

Ruas, J. L. ^{[3
]}

Farias, N. A. R. ^{[2
]}

Rev-Valeiron, C. ^{[4
]}

Pingret, U. ^{[5
]}

Etievant, M. ^{[5
]}

Barba-Carretero, J. C. ^{[6
]}

机构：

[1] Univ Alcala de Henares, Fac Farm, Dept Microbiol & Parasitol, Alcala De Henares 28871, Spain

[2] Univ Fed Pelotas, Inst Biol, Dept Microbiol & Parasitol, Pelotas, Brazil

[3] Univ Fed Pelotas, Fac Vet LRD, Pelotas, Brazil

[4] Univ Nacl Expt Francisco Miranda, Program Ciencias Vet, Dept Sanidad Anim, Falcon, Venezuela

[5] Scanelis ENVT, Toulouse, France

[6] Lab Diagnost Vet, Madrid 28005, Spain

来源：

VETERINARY PARASITOLOGY | 2007年 / 150卷 / 04期

关键词：

Hepatozoon sp; quantitative PCR; dog; cat;

D O I：

10.1016/j.vetpar.2007.09.025

中图分类号：

R38 [医学寄生虫学]; Q [生物科学];

学科分类号：

07 ; 0710 ; 09 ; 100103 ;

摘要：

With the aim to improve current molecular diagnostic techniques of Hepatozoon sp. in carnivore mammals, we developed a quantitative PCR (qPCR) assay with SYBR Green I-(R). The method, consisting of amplification of a 235 bp fragment of the 18S rRNA gene, is able to detect at least 0.1 fg of parasite DNA. Reproducible quantitative results were obtained over a range of 0.1 ng 0.1 fg of Hepatozoon sp. DNA. To assess the performance of the qPCR assay, DNA samples from dogs (140) and cats (50) were tested with either standard PCR or qPCR. Positive samples were always confirmed by partial sequencing of the 18S rRNA gene. Quantitative PCR was 15.8% more sensitive than standard PCR to detect H. canis in dogs. In cats, no infections were detected by standard PCR, compared to two positives by qPCR (which were infected by H. canis as shown by sequencing). (c) 2007 Elsevier B.V. All rights reserved.

引用

页码：352 / 356

页数：5

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