RESUMEN
The objective of the study was to develop a bio-safe synthetic peptide ELISA for the detection of antibodies against the infectious bronchitis virus (IBV) using a novel multiple antigenic peptide approach (MAP). After initial ELISA optimization, diagnostic sensitivity (DSn) and specificity (DSp) for the linear peptides were determined using receiver operator curve (ROC) analysis. The peptide IBVP1 showed 90.44% DSn and 88.64% DSp at ROC cut off 22.8% while IBVP2 showed 88.24% DSn and 85.23% DSp at ROC cut off 23.05%. The multimerization of linear peptides to MAP design resulted in the improvement of the diagnostic efficiency up to 94.85% DSn and 92.05% DSp for IBVM1 with 19.95% cut off. A similar improvement in the performance was also observed with 92.65% DSn and 90.91% DSp for IBVM2 at 20.72% cut off. All the peptides were tested for diagnostic specificity and did not show the cross-reactivity with Newcastle disease virus and infectious bursal disease virus positive serum samples. In addition, repeatability testing for all linear and multimeric peptide showed that the coefficient of variation for intra-assay was within the expected limits, ranging from 2.4 to 10.4% and inter-assay coefficient of variation was ranging from 5.56 to 14.3%. In a nutshell, the present study used predicted B cell epitope, the synthetic peptide in linear and multimeric design for IBV antibody detection. The study also highlights peptide antigen with modified scaffold design could be a safe alternative to whole virion-based ELISA for IBV antibody detection.
RESUMEN
BACKGROUND: Biochemical and haematological parameters have not been determined in Bactrian camels kept at high altitude. Therefore, this study was undertaken to characterise different physiological, haematological, biochemical, and morphometric parameters of Bactrian camels of high altitude. For this, total fourteen high altitude healthy Bactrian camels were selected from Leh-Ladakh, India, a high altitude area, and thereafter divided into three age groups (N = 3 young; N = 6 adult; N = 5 old camels) to characterise for above parameters. All the results were compared with Lowlander Bactrian camels. RESULTS: Morphometric measurement showed significant difference in body height, body length, front-hump height and girth, back-hump height and girth, abdomen girth, neck length, and circumference of the shank in the young age group camels as compared to other age groups of Bactrian camels (p < 0.05). Furthermore, all the physiological and haematological parameters were similar in all the age groups of camels (p < 0.05). However, the leukocyte, erythrocyte, Hb, platelets, monocyte, and ESR level were towards the higher side of the normal reference range of Lowlander Bactrian camels. Whereas, the biochemical analysis revealed a significant increase in triglycerides and decrease in protein levels in the younger age group as compared to other age groups (p < 0.05). Although, albumin, aspartate aminotransferase, iron, magnesium, urea, and creatinine levels were insignificant among the different groups, but observed towards the higher side of the low altitude reference range. Interestingly, the glucose levels in all the groups were observed towards the lower side of the range, which showed metabolic adaptation to high altitude. CONCLUSION: These findings suggested there is morphometric and biochemical variation in Bactrian camel of high altitude. The results further helped in establishing novel reference ranges for these parameters in Highlander Bactrian camel. Hence, this study will be the basis of future research on a Bactrian camel from high-altitude cold desert and helpful for better camel husbandry and health management in high altitude.