DR NARESH KUMAR,ALIA KHAN,SHIVANI ARORA,DR FALGUNI PATRA,DR MEENAKSHI DAHIYA,RAGHU H V,DR MANDEEP BALHARA,PRADEEP KUMAR,SULEMAN SHAIKH
申请号：
IN2213/DEL/2014
公开号：
IN2014DE02213A
申请日:
2014.08.05
申请国别(地区):
IN
年份:
2016
代理人:
摘要：
Antibiotics are widely used in dairy cattle management for the treatment of diseases and as dietary supplement. The presence of these antibiotic residues in milk is a serious concern in terms of public health and processing especially fermented milk products. The present invention is based on the markers enzyme (s) released during spore germination in presence of specific analyte i.e. antibiotics. For the production of enzyme-spore based assay (s), B. stearothermophillus strains were screened for different enzymes namely acetyl esterase, esculinase, 6-D-galactosidase, B-D-glucosidase, a-D-galactosidase, a-Dglucosidase and a-D-mannosidase etc. and marker enzymes were selected. This is followed by spore production to an extent of 80-85% in novel "sporulation medium" which primarily contains Beef extract, Peptone, Tryptone and NaCl etc. and subsequently their conversion / germination in "spore germination medium" containing principal components namely Beef extract. Yeast extract. Peptone, Tryptone, Dextrose, Starch and Agar etc. as germinant and nutrient components. A reproducible, real time, simplified enzyme-spore assay (s) in two stages using solid media was developed. An eppendorf tube containing germination medium "seeded with spores was developed based on inhibition by specific analyte i.e. antibiotics in milk and related products. The working principle of enzyme-spore based assay (s) is based on release of specific indicator enzyme (s) by spores during germination which will act specifically on chromogenic substrate resulting in colored reaction end product which is measured semiquantitatively by visual color reaction. The Enzyme-spore based assay (s) exhibits significant correlation with minimal false positive/negative observations at codex recommended concentrations of antibiotics residues when compared with microbial based receptor assay (Charm 11 assay) and ROSA test. The assay showed excellent performance in buffer / or milk system in natural / or spiked sa
