SIMULTANEOUS MULTIPLE DETECTION OF BACTERIAL CAUSES OF BOVINE MASTITIS USING LATERAL FLOW IMMUNO CHROMATOGRAPHIC ASSAY | ||||
| Journal of the Egyptian Veterinary Medical Association | ||||
| Volume 79, Issue 2, June 2019, Page 587-607 PDF (1.14 MB) | ||||
| Document Type: Original Research Articles | ||||
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| Abstract | ||||
| Bovine mastitis continues to be the most economically important affection of dairy cattle. Several microbial agents alone or mixed are incriminated as a causative agent of bovine mastitis. Staphylococcus auras, Streptococcus pyogens, Streptococcus agalactiae, Pseudomonas aeruginosa, K. pneumoniae, E. coli and Candida albicans are among the most commonly isolated pathogens. The conventional bacteriological isolation and identification of these agents are not only time consuming, requiring full equipped laboratory but also it needs experienced personnel. In the present work a lateral flow device (LFD) was developed for simultaneous multiple identification of three of the causative agents of mastitis in few minutes. Rabbit polyclonal antibody (rPAb) specific to the antigenic components of the 3 of the above mentioned microbial agents, namely S. aureus, E. coli, and K. pneumoniae were conjugated with colloidal gold as the detector antibody. It was laid on the conjugate pad. Different Guinea pigs polyclonal antibodies (gPAb) specific to the different antigens tested were used as the capture antibody at the test line (T). Goat anti-rabbit IgG antibody (GAR) was used as the capture antibody at the control line (C) of nitrocellulose strip. The ready-to-use strips were held in a device consists of 3 strip-holding channels each strip for detection of the 3 different bacterial antigens. The minimal microbial counts that gave positive result using the developed LFD were 100CFU/100µl. The sensitivity, specificity and accuracy of LFD for detection of E. coli as compared to bacteriological examination reached to 93.7%, 95.3% and 94.6% respectively. In detection of S. aureusit reached to 82.6%, 92.3% and 89.3%, respectively, while in case of K. pneumoniae it reached to 94.1%, 98.2% and 97.3%, respectively. It has been proved that treatment of the milk samples with a prepared application buffer associated with pre-incubation of tested milk samples in TSB for 6 hr at 37°C increased significantly the sensitivity results of prepared LFD kits. The developed kit proved simple, convenient and results can be obtained in less than 15 min | ||||
| Keywords | ||||
| Bovine mastitis; S. aureus; E. coli; K. pneumoniae; polyclonal antibodies; lateral flow device | ||||
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