Research Article: Evaluation of a commercial Erns-capture ELISA for detection of BVDV in routine diagnostic cattle serum samples

Date Published: March 13, 2007

Publisher: BioMed Central

Author(s): Jaruwan Kampa, Karl Ståhl, Lena HM Renström, Stefan Alenius.

http://doi.org/10.1186/1751-0147-49-7

Abstract

Bovine viral diarrhoea virus (BVDV) is an important pathogen in cattle. The ability of the virus to cross the placenta during early pregnancy can result in the birth of persistently infected (PI) calves. These calves shed the virus during their entire lifespan and are the key transmitters of infection. Consequently, identification (and subsequent removal) of PI animals is necessary to rapidly clear infected herds from the virus. The objective of this study was to evaluate the suitability of a commercial Erns-capture ELISA, in comparison to the indirect immunoperoxidase test (IPX), for routine diagnostic detection of BVDV within a control programme. In addition, the effect of passive immunity and heat-inactivation of the samples on the performance of the ELISA was studied.

In the process of virus clearance within the Swedish BVDV control programme, all calves born in infected herds are tested for virus and antibodies. From such samples, sent in for routine diagnostics to SVA, we selected 220 sera collected from 32 beef herds and 29 dairy herds. All sera were tested for BVDV antigen using the Erns ELISA, and the results were compared to the results from the IPX used within the routine diagnostics.

All 130 samples categorized as virus negative by IPX were tested negative in the ELISA, and all 90 samples categorized as virus positive were tested positive, i.e. the relative sensitivity and specificity of the ELISA was 100% in relation to IPX, and the agreement between the tests was perfect.

We can conclude that the Erns ELISA is a valid alternative that has several advantages compared to IPX. Our results clearly demonstrate that it performs well under Swedish conditions, and that its performance is comparable with the IPX test. It is highly sensitive and specific, can be used for testing of heat-inactivated samples, precolostral testing, and probably to detect PI animals at an earlier age than the IPX.

Partial Text

Bovine viral diarrhoea virus (BVDV) is a widely spread cattle pathogen with a significant economic impact on cattle production [1]. The virus interferes with reproductive and immunological functions and causes subsequent losses due to reproductive disorders and impaired herd performance [2,3]. Based on phylogenetic comparison, the virus can be classified into two genotypes: BVDV-1 and BVDV-2. Whereas BVDV-1 has a world-wide distribution, BVDV-2 appears to be highly prevalent only in North America [4,5] and relatively rare in other continents [6,7].

The identification of PI animals (for subsequent elimination) is an essential element in any BVDV control programme, and depends on accurate diagnostic tests, i.e. tests with high sensitivity and specificity that have been thoroughly evaluated for routine diagnostic use. Moreover, for testing of large series of samples it is desirable that a test is user friendly and allows automation. Even though the IPX test currently used in Sweden has shown to be efficient for detection of PI animals, it is evident that the Erns ELISA have several advantages: it is independent of cell cultures, gives a test result within a few hours and is relatively inexpensive both to establish and run [22]. In addition, our results clearly demonstrate that it performs well under Swedish conditions, i.e. for detection of BVDV-1, and that its performance is comparable with the IPX test. There was a perfect agreement between the results from the two tests, and the separation between COD values from negative and positive samples was good. Out of 220 samples, 219 had COD values either well below or well above the cut-off. However, one sample, considered as virus positive according to the IPX test results, had a COD value close to the cut-off, and there are a number of possible explanations for this result. Firstly, as with the majority of BVDV antigen ELISAs, this Erns ELISA has been developed for the identification of PI animals. Whereas virus titres in PI animals normally range between 102.2 and 106 TCID50/ml, titres during transient infections have been reported to be as low as 100.9 [23-25]. It is likely that the detection level of the IPX test is lower than that of the ELISA, and it is possible that this serum sample originated from a transiently infected animal. Secondly, although PI animals normally have high virus titres, these may, as previously mentioned, show a wide range. Consequently, it is also possible that this serum sample originated from a PI animal, but that the virus titer was low, and close to the detection limit of the ELISA.

Based on these results we can conclude that the Erns ELISA is a valid alternative to the IPX test. It is highly sensitive and specific, can be used for testing of heat-inactivated samples, precolostral testing, and probably to detect PI animals at an earlier age than the IPX. However, it should be kept in mind that this ELISA, unlike the IPX, uses MAbs, and that it therefore is less likely to detect atypical pestivirus strains.

The author(s) declare that they have no competing interests.

JK and SA took part in all aspects of the study, including study design, laboratory analysis, interpretation of the results, and drafting of the manuscript. KS participated in interpretation of the results and drafting of the manuscript. LR participated in study design and revision of the manuscript. All authors have read and given final approval of the version to be published.

 

Source:

http://doi.org/10.1186/1751-0147-49-7

 

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