Evaluation of ethanol vortex ELISA for detection of bovine tuberculosis in cattle and deer

Abstract

BackgroundThe use of serological assays for diagnosis of bovine tuberculosis (TB) has been intensively studied and use of specific antigens have aided in improving the diagnostic accuracy of the assays. In the present study, we report an in-house enzyme linked immunosorbent assay (ELISA), developed by using ethanol extract of Mycobacterium bovis (M. bovis). The assay, named (ethanol vortex ELISA [EVELISA]), was evaluated for detection of anti- M. bovis antibodies in the sera of cattle and white-tailed deer.MethodsBy using the EVELISA, we tested sera obtained from two species of animals; cattle (n = 62 [uninfected, n = 40; naturally infected, n = 22]) and white-tailed deer (n = 41 [uninfected, n = 25; naturally infected, n = 7; experimentally infected, n = 9]). To detect species specific molecules, components in the ethanol extract were analyzed by thin layer chromatography and western blotting.ResultsAmong the tested animals, 77.2% of infected cattle and 87.5% of infected deer tested positive for anti- M. bovis antibody. There were only minor false positive reactions (7.5% in cattle and 0% in deer) in uninfected animals. M. bovis -specific lipids and protein (MPB83) in the ethanol extract were detected by thin layer chromatography and western blotting, respectively.ConclusionThe results warrant further evaluation and validation of EVELISA for bovine TB diagnosis of traditional and alternative livestock as well as for free-ranging animal species.

Characterization of Ethanol Extracted Cell Wall Components of Mycobacterium avium Subsp. paratuberculosis

“…Surprisingly, the 35 kDa major membrane protein was not present in this extract, as confirmed by the lack of reactivity with two mAbs to independent epitopes within this protein [23]. The MPB83 protein from M. bovis has long been reported as a surface-exposed lipoglycoprotein in M. bovis [24,25]; this protein is present in EtOH extracts, as demonstrated previously [4]. MPB83 was confirmed in M. bovis EtOH extracts at the expected size of 22-kDa in this study (Figure 7b), although it does not correspond to the most abundant protein present in these extracts.…”

The prevalence estimates of Mycobacterium bovis infection in cattle with ELISA

“…Thus, it makes favorable to use tests which are able to detect antibody response. The ELISA technique that has been applied for the diagnosis of bovine tuberculosis and claimed extremely advantageous to identify infected cows, enables their separation from the herd and assists disease eradication (Lilenbaum L and Fonseca, 2006;Wadhwa et al, 2014). This study was conducted in adult cattle, all were over the age of 5 years, attempting to detect the animals in advanced phase of disease through the specific antibody response as the final outcome of infection.…”

Prevalence and associated risk factors for Mycobacterium avium subsp. paratuberculosis in dairy cattle in Mexico

“…The ability to rapidly diagnose and identify the causative agent are critical for combating diseases and stopping epidemics (Wadhwa et al, 2012;Kaur et al, 2013). Recent technological developments have led to the proliferation of new and improved diagnostic tests that hold promise for a better management and control of infectious diseases (Wadhwa et al, 2014). New technologies such as microfluidics (Wadhwa et al, 2012) and "Lab-on-Chip" (Liu et al, 2011) are examples of promising new technologies with the potential to be used as a laboratory-free diagnostic devices for infectious diseases in animal husbandry.…”