Evaluation of the sensitivity of PCR methods for the detection of extraneous agents and comparison with in vivo testing
Evaluation of the sensitivity of PCR methods for the detection of extraneous agents and comparison with in vivo testing
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DOI：
10.1016/j.biologicals.2010.01.002
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发表时间：
2010-05-01
期刊：
影响因子：
1.7
通讯作者：
Jungbaeck, C.
中科院分区：
文献类型：
--
作者：
Motitschke, A.;Ottiger, H. P.;Jungbaeck, C.
本研究比较了聚合酶链反应（PCR）方法检测新城疫病毒（NDV）、禽呼肠孤病毒（ARV）、禽流感病毒（AIV）和禽传染性支气管炎病毒（IBV）的敏感性与欧洲药典（Ph. Eur.）中相应血清学检测方法的敏感性。为此，分别配制了10倍稀释的灭活疫苗，并给SPF级鸡组接种了两倍稀释的疫苗。21天后，这些动物再次接种单剂量疫苗。两周后，使用Idexx酶联免疫吸附试验（ELISA）对每只动物的血清样本进行抗体检测。同时，对稀释后的疫苗样本进行PCR检测。结果发现，四种PCR检测的灵敏度与相应的血清学检测相当，甚至略好于血清学检测。因此，这些聚合酶链反应试验满足Ph. Eur的灵敏度要求。并且可以用作检测最后一批灭活疫苗中外来剂的替代测试。(C) 2010国际生物制品协会。Elsevier Ltd.出版。版权所有。
In this study the sensitivity of polymerase chain reaction (PCR) methods for the detection of Newcastle disease virus (NDV), avian reovirus (ARV), avian influenza virus (AIV) and avian infectious bronchitis virus (IBV) was compared to the sensitivity of the corresponding serological tests described in the European Pharmacopoeia (Ph. Eur.). For this purpose, serial 10-fold dilutions of the respective inactivated vaccines were prepared and groups of SPF chickens were vaccinated with a double dose of the vaccine dilutions. After a period of 21 days, the animals were revaccinated with a single dose. Two weeks later, serum samples from each animal were tested for antibodies using an Idexx enzyme linked immunosorbent assay (ELISA). In parallel, samples of the diluted vaccines were tested by PCR. It was found that the sensitivity of the four PCR tests is comparable to or even slightly better than that of the corresponding serological tests. Thus these PCR tests fulfil the sensitivity requirements of the Ph. Eur. and could be used as alternative tests for the detection of extraneous agents in final batches of inactivated vaccines. (C) 2010 The International Association for Biologicals. Published by Elsevier Ltd. All rights reserved.