High throughput Luminex beads based multiplex assay for identification of six major bacterial pathogens of mastitis in dairy animals
IntroductionBovine mastitis is caused by over 150 different microorganisms. Specific identification and quantification of multiple bacteria in a single milk sample becomes essential for rapid intervention.MethodsIn the present study a Luminex beads based multiplex assay emphasizing on the precise id...
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Frontiers Media S.A.
2023-07-01
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| Series: | Frontiers in Cellular and Infection Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fcimb.2023.1125562/full |
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| author | Garima Shrinet Rajesh Chhabra Archana Sharma Kanisht Batra Saurabh Jyoti Talukdar Sushila Maan |
| author_facet | Garima Shrinet Rajesh Chhabra Archana Sharma Kanisht Batra Saurabh Jyoti Talukdar Sushila Maan |
| author_sort | Garima Shrinet |
| collection | DOAJ |
| description | IntroductionBovine mastitis is caused by over 150 different microorganisms. Specific identification and quantification of multiple bacteria in a single milk sample becomes essential for rapid intervention.MethodsIn the present study a Luminex beads based multiplex assay emphasizing on the precise identification of six major bacterial pathogens of mastitis was developed. Assay was developed in two triplex sets, triplex 1 comprised of Streptococcus agalactiae, Streptococcus dysgalactiae and Streptococcus uberis while triplex 2 consisted of Staphylococcus aureus, E. coli and Klebsiella pneumoniae.ResultsThe analytical sensitivity was 10 6 copies per reaction mixture for all the six bacteria. A 100% analytical specificity was observed for simultaneous detection of these bacteria. Clinical milk samples from 100 bovine quarters were tested for validation.DiscussionThe analytical sensitivity was similar to the findings reported earlier in real time PCR multiplex assay targeting the DNA of the 11 most common bacterial species or groups in mastitis. The analytical specificity of the optimized assay was 100% similar to reported earlier for simultaneous detection of Mycoplasma spp. and for seven entric viruses of humans.The developed assay indicates a concept proof of a rapid, cost effective high throughput diagnostic tool for identification of major bacteria causing mastitis. |
| first_indexed | 2024-03-12T23:08:28Z |
| format | Article |
| id | doaj.art-4619164bbced4a2fb6160f08bcb82a54 |
| institution | Directory Open Access Journal |
| issn | 2235-2988 |
| language | English |
| last_indexed | 2024-03-12T23:08:28Z |
| publishDate | 2023-07-01 |
| publisher | Frontiers Media S.A. |
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| series | Frontiers in Cellular and Infection Microbiology |
| spelling | doaj.art-4619164bbced4a2fb6160f08bcb82a542023-07-18T08:49:14ZengFrontiers Media S.A.Frontiers in Cellular and Infection Microbiology2235-29882023-07-011310.3389/fcimb.2023.11255621125562High throughput Luminex beads based multiplex assay for identification of six major bacterial pathogens of mastitis in dairy animalsGarima Shrinet0Rajesh Chhabra1Archana Sharma2Kanisht Batra3Saurabh Jyoti Talukdar4Sushila Maan5Department of Veterinary Microbiology, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, Haryana, IndiaCollege Central Laboratory, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, Haryana, IndiaDepartment of Veterinary Microbiology, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, Haryana, IndiaDepartment of Animal Biotechnology, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, Haryana, IndiaCollege Central Laboratory, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, Haryana, IndiaDepartment of Animal Biotechnology, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, Haryana, IndiaIntroductionBovine mastitis is caused by over 150 different microorganisms. Specific identification and quantification of multiple bacteria in a single milk sample becomes essential for rapid intervention.MethodsIn the present study a Luminex beads based multiplex assay emphasizing on the precise identification of six major bacterial pathogens of mastitis was developed. Assay was developed in two triplex sets, triplex 1 comprised of Streptococcus agalactiae, Streptococcus dysgalactiae and Streptococcus uberis while triplex 2 consisted of Staphylococcus aureus, E. coli and Klebsiella pneumoniae.ResultsThe analytical sensitivity was 10 6 copies per reaction mixture for all the six bacteria. A 100% analytical specificity was observed for simultaneous detection of these bacteria. Clinical milk samples from 100 bovine quarters were tested for validation.DiscussionThe analytical sensitivity was similar to the findings reported earlier in real time PCR multiplex assay targeting the DNA of the 11 most common bacterial species or groups in mastitis. The analytical specificity of the optimized assay was 100% similar to reported earlier for simultaneous detection of Mycoplasma spp. and for seven entric viruses of humans.The developed assay indicates a concept proof of a rapid, cost effective high throughput diagnostic tool for identification of major bacteria causing mastitis.https://www.frontiersin.org/articles/10.3389/fcimb.2023.1125562/fullLuminexmastitisbovinebacteriahigh throughputdiagnosis |
| spellingShingle | Garima Shrinet Rajesh Chhabra Archana Sharma Kanisht Batra Saurabh Jyoti Talukdar Sushila Maan High throughput Luminex beads based multiplex assay for identification of six major bacterial pathogens of mastitis in dairy animals Frontiers in Cellular and Infection Microbiology Luminex mastitis bovine bacteria high throughput diagnosis |
| title | High throughput Luminex beads based multiplex assay for identification of six major bacterial pathogens of mastitis in dairy animals |
| title_full | High throughput Luminex beads based multiplex assay for identification of six major bacterial pathogens of mastitis in dairy animals |
| title_fullStr | High throughput Luminex beads based multiplex assay for identification of six major bacterial pathogens of mastitis in dairy animals |
| title_full_unstemmed | High throughput Luminex beads based multiplex assay for identification of six major bacterial pathogens of mastitis in dairy animals |
| title_short | High throughput Luminex beads based multiplex assay for identification of six major bacterial pathogens of mastitis in dairy animals |
| title_sort | high throughput luminex beads based multiplex assay for identification of six major bacterial pathogens of mastitis in dairy animals |
| topic | Luminex mastitis bovine bacteria high throughput diagnosis |
| url | https://www.frontiersin.org/articles/10.3389/fcimb.2023.1125562/full |
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