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基于ITS1 rDNA的聚合酶链反应在艾美耳球虫分类鉴定中的应用

Application of polymerase chain reaction based on ITS1 rDNA to speciate Eimeria.

作者信息

Jenkins M C, Miska K, Klopp S

机构信息

Animal Parasitic Diseases Laboratory, Beltsville Agricultural Research Center, Agricultural Research Service, USDA, Beltsville, MD 20705, USA.

出版信息

Avian Dis. 2006 Mar;50(1):110-4. doi: 10.1637/7439-091305R.1.

Abstract

A method was developed to recover Eimeria spp. oocysts directly from poultry litter and determine which species of Eimeria were present using polymerase chain reaction (PCR) based on the ITS1 rDNA sequence. The species composition of Eimeria oocysts was also compared before and after propagation in susceptible chickens to determine if the relative proportion of each species changed after expansion. In samples from two broiler operations, ITS1-PCR was able to detect Eimeria spp. oocysts recovered from litter, with Eimeria acervulina, Eimeria maxima, and Eimeria praecox being the predominant species present therein. Although Eimeria tenella was found in one sample, the other species--Eimeria brunetti, Eimeria necatrix, and Eimeria mitis-were not detected. The species composition as determined by ITS1-PCR did not appear to appreciably alter after expansion in susceptible chickens. The described method represents a rapid means for determining the major Eimeria species in a poultry operation and may be helpful in choosing a particular live oocyst vaccine formulation to protect chickens against coccidiosis.

摘要

开发了一种直接从家禽粪便中回收艾美耳球虫属卵囊的方法,并基于ITS1 rDNA序列使用聚合酶链反应(PCR)来确定存在哪些艾美耳球虫种类。还比较了在易感鸡中传代前后艾美耳球虫卵囊的种类组成,以确定每种种类在扩增后相对比例是否发生变化。在来自两个肉鸡养殖场的样本中,ITS1-PCR能够检测到从粪便中回收的艾美耳球虫属卵囊,其中堆型艾美耳球虫、巨型艾美耳球虫和早熟艾美耳球虫为主要种类。虽然在一个样本中发现了柔嫩艾美耳球虫,但未检测到其他种类——布氏艾美耳球虫、毒害艾美耳球虫和微小艾美耳球虫。经ITS1-PCR测定的种类组成在易感鸡中扩增后似乎没有明显改变。所描述的方法是确定家禽养殖场中主要艾美耳球虫种类的快速手段,可能有助于选择特定的活卵囊疫苗制剂来保护鸡免受球虫病感染。

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