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#28752213   2017/07/28 Save this To Up

Changes of haemogram and serum biochemistry in neonatal piglet diarrhoea associated with porcine rotavirus type A.

Porcine rotavirus type A (RVA) is a major cause of neonatal piglet mortality in India. The effect of the disease on haemogram and serum biochemical profile is not well established in piglets. Accordingly, we assessed the haemogram and serum biochemical profile in the neonatal piglet diarrhoea with RVA infection (n = 17). The diagnosis of RVA was confirmed using RNA-polyacrylamide gel electrophoresis (RNA-PAGE), commercially available enzyme-linked immunosorbent assay (ELISA) kit and reverse transcription-polymerase chain reaction (RT-PCR). Non-infected healthy piglets (n = 6) served as control. The concentrations of total protein, albumin, alanine amino transaminase (ALT), aspartate amino transaminase (AST), blood urea nitrogen (BUN) and creatinine in serum were measured by spectrophotometric method. Haemogram was done in the blood using sodium ethylenediaminetetraacetic acid (Na2 EDTA) as anticoagulant. The mean values of total protein, albumin and globulin concentrations were significantly (P < 0.001) decreased and concentrations of ALT, AST, BUN and creatinine were significantly increased (P < 0.001) in the RVA-infected piglets. Haemogram showed marked haemoconcentration (P < 0.001), leukopenia (P < 0.01) and neutropenia (P < 0.01) in the presence of RVA infection than healthy piglets. The results indicated a possible extra-intestinal spread of RVA in piglets during neonatal diarrhoea. The finding might be helpful to clinicians and while treating such type of clinical cases, incorporation of organ protective drugs will be helpful for better response in the treatment schedule.

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#23953456   2013/08/19 Save this To Up

Ru(phen)3(2+) doped silica nanoparticle based immunochromatographic strip for rapid quantitative detection of β-agonist residues in swine urine.

A Ru(phen)3(2+) doped silica nanoparticle based immunochromatographic strip was developed for the rapid and quantitative detection of five common β-agonist (salbutamol (SAL), cimbuterol, terbutaline, clenbuterol, and brombuterol) residues in swine urine. The broad spectrum monoclonal antibodies generated by immunizing BALB/c mice with salbutamol conjugated cationic bovine serum albumin. The fluorescence intensities (FIs) of the strip on the test line (FIT) and control line (FIC) were determined using a strip reader. Parameters that influenced the antibody and antigen interaction on the test strip were investigated by recording FIT and FIC values, and the concept of FIT/FIC ratio was used to offset the inherent heterogeneity of the test strips and the effect of the sample matrix. Under optimal conditions, the linear range for the quantitative detection of SAL was 0.6-5.0 ng/ml with a half maximal inhibitory concentration at 1.78 ng/ml. The limit of detection for real swine urine was 0.43 ng/ml. The recovery rates of the intraassay for spiked urine at SAL concentrations of 0.8, 1.5, and 3.5 ng/mL were 88.06%±3.75%, 95.77%±5.33%, and 94.06%±7.43%, whereas those for the interassay were 84.69%±5.0%, 95.06%±9.3%, and 88.34%±7.71%, respectively. The developed quantitative method exhibited excellent agreement with a commercially available competitive enzyme-linked immunosorbent assay kit for SAL-spiked urine samples, with a correlation of coefficient of 0.95 and a slope of 0.99 (n=36). The results indicated that the developed test strip enables sensitive, reproducible, and easily implementable screening for the rapid and quantitative detection of β-agonist residues in swine urine.

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#19236874   2009/04/13 Save this To Up

A robust quantitative solid phase immunoassay for the acute phase protein C-reactive protein (CRP) based on cytidine 5'-diphosphocholine coupled dendrimers.

C-reactive protein (CRP) is an important acute phase protein, being used as a sensitive indicator of inflammation and infection and is also associated with the risk of cardiovascular problems. The present paper describes a robust and sensitive ELISA for CRP, based on the affinity of CRP for phosphocholine. In this design synthetic globular polymers (dendrimers) are used as scaffolds for the multivalent display of phosphocholine molecules. CRP present in a sample binds to the phosphocholine moiety presented at high density in the coating layer and is detectable by specific antibodies. The ELISA was applied to determination of pig and human CRP using commercially available antibodies against human CRP. The assay was shown to be more sensitive than previously published immunoassays employing albumin-coupled cytidine diphosphocholine. The coating was stable for at least 30 days at room temperature and the assay showed high intra- and interassay reproducibility. Results were compared with an immunoturbidimetric method and with a commercial ELISA kit and there was very good agreement with the immunoturbidimetric method, however not with the commercial assay, probably due to a calibration discrepancy. The assay is applicable to other species by providing an adequate detection antibody having the desired species specificity.

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#1403424   1992/11/10 Save this To Up

Minimizing ELISA background in the diagnosis of swine trichinellosis.

After confirming that long-term serum storage (frozen at -20 C for greater than 3 mo) causes optical density to drift upward, several modifications of an enzyme-linked immunosorbent assay (ELISA) protocol were evaluated to identify a protocol that would reduce background in porcine sera tested for trichinellosis. Modifications evaluated included blocking the antigen-coated ELISA plate with sample diluent containing 10% bovine serum albumin (BSA) or 10% nonfat milk powder (bovine lacto transfer optimizer or BLOTTO), diluting sera in sample diluent containing 10% BSA or 10% BLOTTO, and preincubating samples in sample diluent containing 10% BSA or 10% BLOTTO. Overnight preincubation (approximately 12 hr at 2 C) of fresh sera diluted (1:10) in sample diluent containing 10% BLOTTO significantly reduced background and improved the detection of experimentally infected pigs by enhancing positive-negative discrimination. When testing stored sera, the modified protocol effectively reduced the effect of storage and the kit revealed specificity of 98.4%; there was no loss in sensitivity. The effect of long-term storage at -20 C must therefore be considered when testing swine sera for trichinellosis by ELISA and possibly also when conducting other immunoglobulin assays. The modification described here may prove useful if there is no alternative to using serum stored for greater than 3 mo at -20 C.

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