Oral fluid analysis for herd monitoring is of interest to the commercial pig production in Korea. The aim of this study was to investigate pathogen-positive rates and correlations among eight pathogens associated with porcine respiratory disease complex by analyzing oral fluid samples from 214 pig groups from 56 commercial farms. Samples collected by a rope-chewing method underwent reverse-transcriptase polymerase chain reaction (RT-PCR) or standard polymerase chain reaction (PCR) analysis, depending on the microorganism. Pathogens were divided into virus and bacteria groups. The former consisted of porcine reproductive and respiratory syndrome virus and porcine circovirus type 2 (PCV2), and the latter Pasteurella multocida, Haemophilus parasuis, Actinobacillus pleuropneumoniae, Mycoplasma hyopneumoniae (MHP), Mycoplasma hyorhinis, and Streptococcus suis (SS). All pathogens were detected more than once by PCR. Age-based analysis showed the PCR-positive rate increased with increasing age for PCV2 and MHP, whereas SS showed the opposite. Correlations between pathogens were assessed among 36 different pair combinations; only seven pairs showed statistically significant correlations. In conclusion, the oral fluid method could be a feasible way to detect various swine respiratory disease pathogens and, therefore, could complement current monitoring systems for respiratory diseases in the swine industry.
BackgroundFoot-and-mouth disease (FMD) can be controlled by either stamping out or vaccination, a choice which depends on both the economic importance of the livestock sector as well as the disease status. In FMD-free countries with vaccination, such as Korea, vaccination programs should guarantee prevention against transmission of FMD. Monitoring of vaccination programs is also essential for ensuring sufficient coverage that will limit the transmission of FMDV. There are several methods to screen FMD virus (FMDV) structural protein (SP) antibodies including SPCE (Solid-phase competitive ELISA), LPBE (Liquid-phase blocking ELISA), and VNT (Virus neutralization test). Among these, SPCE is widely used for serological monitoring since VNT—the gold standard method—has certain practical limitations, such as high costs in terms of time and labor. However, whether SPCE can ensure the vaccination status of individual animals and whole farms is unclear. In this study, SPCE, LPBE and VNT were compared with respect to correlation with each other and sensitivity at commercial pig farms.ResultsThe positive results obtained by PrioCHECK SPCE differed from those obtained by LPBE and VNT. The sensitivity of SPCE relative to those of the other tests was fairly low. The raw data of SPCE were most highly correlated with those of VNT with XJ strain, while their positivity and negativity were most highly correlated with LPBE. The results of ROC analysis proposed new cut-off for PrioCHECK SPCE higher than the previous 50% inhibition.ConclusionsThe high false positive rate of PrioCHECK SPCE suggested that high seropositivity by SPCE may not guarantee a true vaccination coverage. Adjusting the cut-off percentage (%) inhibition value for SPCE is needed to address this problem, and it is highly recommended that routine FMDV serological monitoring programs using PrioCHECK SPCE should be combined with alternative methods such as LPBE or VNT.
Simple methods for measuring the levels of serum antibody against canine distemper virus (CDV) would assist in the effective vaccination of dogs. To develop an enzyme-linked immunosorbent assay (ELISA) specific for CDV, we expressed hydrophilic extra-viral domain (HEVD) protein of the A75/17-CDV H gene in a pET 28a plasmid-based Escherichia (E.) coli vector system. Expression was confirmed by dot and Western blotting. We proposed that detection of E. coli-expressed H protein might be conformation-dependent because intensities of the reactions observed with these two methods varied. The H gene HEVD protein was further purified and used as an antigen for an ELISA. Samples from dogs with undetectable to high anti-CDV antibody titers were analyzed using this HEVD-specific ELISA and a commercial CDV antibody detection kit (ImmunoComb). Levels of HEVD antigenicity measured with the assays and immunochromatography correlated. These data indicated that the HEDV protein may be used as antigen to develop techniques for detecting antibodies against CDV.
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