Development and validation of a loop mediated isothermal amplification (LAMP) assay for the detection of Staphylococcus aureus in bovine mastitis milk samples

Sheet, Omar H.; Grabowski, Nils Th.; Klein, G.; Abdulmawjood, Amir

doi:10.1016/j.mcp.2016.08.001

Cited by 66 publications

(34 citation statements)

References 25 publications

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“…A relatively novel mastitis-diagnostic with potential for short time-to-result is loop-mediated isothermal amplification (LAMP). LAMP primers are available for S. aureus (Sheet et al, 2016) and major streptococci (Bosward et al, 2016;Wang & Liu, 2015), and implementation as pregnancy-test-like lateral flow device is possible (Cornelissen et al, 2016). A major challenge for on-farm molecular detection of pathogens or AMR genes is the large number of bacterial species and resistance genes that may be present in milk or mastitis pathogens.…”

Section: Detectionmentioning

confidence: 99%

An update on environmental mastitis: Challenging perceptions

Klaas

Zadoks

2017

Transbound Emerg Dis

187

144

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Section: Detectionmentioning

confidence: 99%

An update on environmental mastitis: Challenging perceptions

Klaas

Zadoks

2017

Transbound Emerg Dis

187

144

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“…The LAMP method is already validated for various topics in many research areas. In the field of food safety, bacterial pathogens [ 10 ] and fungal contaminants [ 11 ] were detected and in case of urgent human health issues, evidence of emerging pathogens like tuberculosis [ 12 ] or zika virus disease [ 13 ] were possible. Furthermore, identifying animal species in meat products, such as ostrich or pork, was conducted with the LAMP technology [ 14 , 15 ].…”

Section: Introductionmentioning

confidence: 99%

Loop-mediated isothermal amplification (LAMP) assay—A rapid detection tool for identifying red fox (Vulpes vulpes) DNA in the carcasses of harbour porpoises (Phocoena phocoena)

et al. 2017

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Carcasses of wild animals are often visited by different scavengers. However, determining which scavenger caused certain types of bite marks is particularly difficult and knowledge thereof is lacking. Therefore, a loop-mediated isothermal amplification (LAMP) assay (target sequence cytochrome b) was developed to detect red fox DNA in carcasses of harbour porpoises. The MSwab™ method for direct testing without prior DNA isolation was validated. As a detection device, the portable real-time fluorometer Genie® II was used, which yields rapid results and can be used in field studies without huge laboratory equipment. In addition to in vitro evaluation and validation, a stranded and scavenged harbour porpoise carcass was successfully examined for red fox DNA residues. The developed LAMP method is a valuable diagnostic tool for confirming presumable red fox bite wounds in harbour porpoises without further DNA isolation steps.

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“…Two extra primers, LF (loop forward) and LB (loop backward), are also incorporated in order to accelerate the amplification of reaction as well as enhance the specificity [10]. LAMP has been shown to be a sensitive and specific method for the detection of veterinary pathogens [11][12][13][14]. The detection of M. agalactiae by LAMP was first reported by Rekha et al [15].…”

Section: Introductionmentioning

confidence: 99%

Validation of Loop-Mediated Isothermal Amplification (LAMP) Field Tool for Rapid and Sensitive Diagnosis of Contagious Agalactia in Small Ruminants

Tumino

Tolone

Parco

et al. 2020

Animals

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Contagious agalactia (CA), an infectious disease of small ruminants, caused by Mycoplasma agalactiae, is responsible for severe losses to dairy sheep production with substantial socioeconomic impacts on small-scale farmers. The diagnosis of CA is still problematic, time-consuming and requires well-equipped labs for confirmation of outbreaks. Therefore, rapid, accurate and cost-effective diagnostic tests are urgently needed. This work aims to validate a novel Loop-Mediated Isothermal Amplification (LAMP) test, based on the p40 target gene, for the detection of M. agalactiae in dairy sheep in order to confirm its potential practical use as a rapid and cheap field test. The LAMP system proposed in this study consists of a portable device composed of real-time fluorometer with the automatic interpretation of results displayed in a tablet. A total of 110 milk samples (90 positives and 20 negatives) were analysed to optimise the analysis procedure and to investigate the efficacy and robustness of the LAMP method. All samples were analysed using LAMP and conventional real-time PCR to compare the diagnostic sensitivity of the methods. The sensitivity of the LAMP was 10-fold higher than that of real-time PCR, with a detection limit up to 103 CFU/ml. The LAMP assay was able to detect M. agalactiae in 81 of 90 (90%, 95%CI 0.84–0.96) positive milk samples compared to 69 (77%, 95%CI 0.59–0.95) positive samples detected by real-time PCR; no positive signal occurred for any of the negative milk samples in either test. Therefore, the LAMP assay was found to be more sensitive than real-time PCR, low-cost, easy to perform, fast and not affected by contamination, indicating its potential as an effective diagnostic tool in the field level for the diagnosis of CA.

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Development and validation of a loop mediated isothermal amplification (LAMP) assay for the detection of Staphylococcus aureus in bovine mastitis milk samples

Cited by 66 publications

References 25 publications

An update on environmental mastitis: Challenging perceptions

An update on environmental mastitis: Challenging perceptions

Loop-mediated isothermal amplification (LAMP) assay—A rapid detection tool for identifying red fox (Vulpes vulpes) DNA in the carcasses of harbour porpoises (Phocoena phocoena)

Validation of Loop-Mediated Isothermal Amplification (LAMP) Field Tool for Rapid and Sensitive Diagnosis of Contagious Agalactia in Small Ruminants

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