Development of a loop-mediated isothermal amplification assay for the detection of Streptococcus agalactiae in bovine milk

Journal of Dairy Science
Katrina L BoswardPaul A Sheehy

Abstract

Streptococcus agalactiae is a well-characterized bovine mastitis pathogen that is known to be highly contagious and capable of spreading rapidly in affected dairy herds. Loop-mediated isothermal amplification (LAMP) is a novel molecular diagnostic method that has the capability to provide rapid, cost-effective screening for pathogens to support on-farm disease control and eradication programs. In the current study, a LAMP test was developed to detect S. agalactiae in milk. The assay was validated on a bank of existing clinical mastitis milk samples that had previously been identified as S. agalactiae positive via traditional microbiological culture techniques and PCR. The LAMP assay was conducted on bacterial colonies and DNA extracted from milk in tube- and plate-based formats using multiple detection platforms. The 1-h assay conducted at 64 °C exhibited repeatability (coefficient of variation) of 2.07% (tube) and 8.3% (plate), sensitivity to ~20 pg of extracted DNA/reaction, and specificity against a panel of known bacterial mastitis pathogens. Of the 109 known S. agalactiae isolates assessed by LAMP directly from bacterial cells in culture, 108 were identified as positive, in accordance with PCR analysis. The LAMP analysis f...Continue Reading

References

Feb 1, 1979·Zentralblatt für Veterinärmedizin. Reihe B. Journal of veterinary medicine. Series B·E Y Bashandy, L E Heider
Jul 15, 1989·The Veterinary Record·Y H SchukkenA Brand
Apr 1, 1972·Journal of Dairy Science·L O LuedeckeJ K Hillers
Jun 28, 2000·Nucleic Acids Research·T NotomiT Hase
Aug 31, 2002·Journal of Dairy Science·I Meiri-BendekY Kashi
Oct 3, 2006·Journal of Biochemical and Biophysical Methods·Hisatoshi KanekoTatsuo Suzutani
Feb 27, 2007·The Veterinary Record·A J BradleyM J Green
Oct 13, 2007·The Journal of Dairy Research·Sofie PiepersSarne De Vliegher
Apr 28, 2009·Journal of Infection and Chemotherapy : Official Journal of the Japan Society of Chemotherapy·Yasuyoshi Mori, Tsugunori Notomi
Feb 9, 2011·Acta Veterinaria Scandinavica·Piret KalmusKalle Kask
Feb 12, 2011·Nucleic Acids Research·Yasumasa KimuraYoshihide Hayashizaki
Feb 14, 2012·Journal of Virological Methods·Adam J FoordHans G Heine
Jun 6, 2012·The Veterinary Clinics of North America. Food Animal Practice·Greg Keefe
Oct 30, 2012·The Veterinary Clinics of North America. Food Animal Practice·Abhijit GurjarPaolo Moroni
Dec 3, 2013·The Veterinary Record·Martin Green, Andrew Bradley

❮ Previous
Next ❯

Citations

Jul 4, 2016·Journal of Dairy Science·Olivier SamsonRuth Zadoks
Aug 25, 2011·Research in Veterinary Science·Acácia Orieth EliasHelio Langoni
Jun 14, 2017·Animal Health Research Reviews·Thabiso E MotaungToi J Tsilo
Oct 31, 2017·Transboundary and Emerging Diseases·I C Klaas, R N Zadoks
Jan 13, 2019·Letters in Applied Microbiology·G SharmaS Sethi
Sep 27, 2018·International Journal of Veterinary Science and Medicine·Amr El-SayedHugo Castañeda Vázquez
Jun 28, 2018·Tropical Animal Health and Production·Aqeela Ashraf, Muhammad Imran
Aug 17, 2019·Frontiers in Bioengineering and Biotechnology·Sofia A M MartinsPaulo P Freitas
Nov 21, 2020·The Journal of Dairy Research·Francisco B MalcataLorenzo Viora
Jun 20, 2018·Microbiology Spectrum·David H Lloyd, Stephen W Page

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.