On-Site Detection of Tissues of Buffalo Origin by Loop-Mediated Isothermal Amplification (LAMP) Assay Targeting Mitochondrial Gene Sequences

Kumari, Sarita; Kumar, Rajiv; Mendiratta, S. K.; Kumar, Deepak; Kumar, Arun; Kumar, Dinesh; Rana, Preeti; Jawla, Jyoti

doi:10.1007/s12161-020-01718-2

Cited by 11 publications

(14 citation statements)

References 30 publications

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“…In the present study, the AL DNA extraction method is combined with LAMP assay to develop a user‐friendly point of care technique for the identification of buffalo meat. LAMP is an isothermal DNA amplification method that has been successfully applied by several authors for meat authentication (Girish et al, 2020; Notomi et al, 2000; Sul, Kim, & Kim, 2019; Yang et al, 2014) including buffalo meat (Kumari et al, 2020). The LAMP method described by Kumari et al (2020) for buffalo meat identification included a commercial kit for DNA extraction thereby making the analysis expensive.…”

Section: Discussionmentioning

confidence: 99%

“…LAMP is an isothermal DNA amplification method that has been successfully applied by several authors for meat authentication (Girish et al, 2020; Notomi et al, 2000; Sul, Kim, & Kim, 2019; Yang et al, 2014) including buffalo meat (Kumari et al, 2020). The LAMP method described by Kumari et al (2020) for buffalo meat identification included a commercial kit for DNA extraction thereby making the analysis expensive. The genomic DNA extracted from meat by AL method in the present study could be conveniently used for isothermal DNA amplification.…”

Section: Discussionmentioning

confidence: 99%

“…The LAMP method was optimized for various parameters like magnesium ion concentration, annealing temperature and betaine concentration. Magnesium ion concentration greatly affects the amplification efficiency in LAMP where high Mg 2+ concentration results in nonspecific amplification while low Mg 2+ concentration is known to reduce amplification efficiency (Kumari et al, 2020; Tomita et al, 2008). Betaine is a modified amino acid that promotes amplification of difficult to amplify sequences of DNA such as G:C rich regions (Rees et al, 1993).…”

Section: Discussionmentioning

confidence: 99%

“…LAMP based species identification methods have been reported for beef (Cho, Dong, & Cho, 2014), pork (Girish et al, 2020; Yang et al, 2014), duck meat (Shi et al, 2017), buffalo (Kumari et al, 2020), and chicken meat (Ahmed, Hasan, Hossain, Saito, & Tamiya, 2010). Xu et al (2017) developed smart and sealed biosensor combining LAMP and lateral flow device (LFD) for meat species identification.…”

Section: Introductionmentioning

confidence: 99%

“…Xu et al (2017) developed smart and sealed biosensor combining LAMP and lateral flow device (LFD) for meat species identification. Kumari et al (2020) developed LAMP for buffalo meat identification using commercial kit for DNA extraction, however, cost of sample analysis is the major concern in such method. Therefore, present study was undertaken to develop a rapid, cost effective, and reliable diagnostic assay for buffalo meat authentication using alkaline lysis (AL) method of DNA extraction and visual detection of amplification by LAMP for on‐field applications that cater to point‐of‐care services.…”

Section: Introductionmentioning

confidence: 99%

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Alkaline lysis‐loop mediated isothermal amplification assay for rapid and on‐site authentication of buffalo (Bubalus bubalis) meat

Girish

Kumari

Gireesh‐Babu

et al. 2021

Journal of Food Safety

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India is the largest producer and exporter of buffalo (Bubalus bubalis) meat in the world. But there are restrictions on the slaughter of cattle (Bos indicus/Bos taurus) and export of beef (cattle meat) from the country. This led to the incidences of misrepresentation of beef and other low cost meat as buffalo meat during export and hence necessitates reliable techniques for species authentication of buffalo meat. Tests need to be rapid to ensure that export consignments are not held for a long time while meat samples are being authenticated. In the present study, a rapid assay for authentication of buffalo meat was developed by employing alkaline lysis (AL) method of DNA extraction and loop‐mediated isothermal amplification (LAMP) using novel primers designed to target mitochondrial D loop region. The results of the AL‐LAMP assay were visualized by SYBR Green I dye. Further, the LAMP amplification was confirmed by agarose gel electrophoresis and fluorometry. The developed AL‐LAMP assay was validated using raw, heat treated, and admixed/adulterated buffalo meat samples. The AL‐LAMP assay could be carried out with minimum instrumentation and hence, holds potential for use as on‐site assay for species authentication of buffalo meat.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Alkaline lysis‐loop mediated isothermal amplification assay for rapid and on‐site authentication of buffalo (Bubalus bubalis) meat

Girish

Kumari

Gireesh‐Babu

et al. 2021

Journal of Food Safety

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Analytical Methods for Authentication of Buffalo Meat

Maheswarappa¹,

Muthukumar²,

Mohan³

et al. 2022

Asiatic Water Buffalo

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Rapid Detection of Hepatitis A Virus in Foods Using a Bioluminescent Assay in Real-Time (BART) and Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) Technology

Meng

Corredig

et al. 2023

Food Environ Virol

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Foodborne hepatitis A infections have been considered as a major threat for public health worldwide. Increased incidences of hepatitis A virus (HAV) infection has been associated with growing global trade of food products. Rapid and sensitive detection of HAV in foods is very essential for investigating the outbreaks. Real-time RT-PCR has been most widely used for the detection of HAV by far. However, the technology relies on fluorescence determination of the amplicon and requires sophisticated, high-cost instruments and trained personnel, limiting its use in low resource settings. In this study, a robust, affordable, and simple assay, reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay in combination with a bioluminescence-based determination of amplification in real-time (BART), was developed for the detection of HAV in different food matrices, including green onion, strawberry, mussel, and milk. The efficiencies of a one-step RT-LAMP-BART and a two-step RT-LAMP-BART were investigated for the detection of HAV in different food matrices and was compared with that of real-time RT-PCR. The sensitivity of the RT-LAMP-BART assay was significantly affected by Mg 2+ concentration ( P < 0.05), in addition to primer quality. The optimal Mg 2+ concentration was 2 mM for one-step RT-LAMP-BART and 4 mM for two-step RT-LAMP-BART. Compared with cartridge-purified primers, HPLC-purified primers could greatly improve the sensitivity of the RT-LAMP-BART assay ( P < 0.05). For detecting HAV in different food matrices, the performance of two-step RT-LAMP-BART was comparable with that of real-time RT-PCR and was better than that of one-step RT-LAMP-BART. The detection limit of the two-step RT-LAMP-BART for HAV in green onion, strawberry, mussel, and milk was 8.3 × 10 0 PFU/15 g, 8.3 × 10 1 PFU/50 g, 8.3 × 10 0 PFU/5 g, and 8.3 × 10 0 PFU/40 mL, respectively. The developed RT-LAMP-BART was an effective, simple, sensitive, and robust method for foodborne HAV detection.

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On-Site Detection of Tissues of Buffalo Origin by Loop-Mediated Isothermal Amplification (LAMP) Assay Targeting Mitochondrial Gene Sequences

Cited by 11 publications

References 30 publications

Alkaline lysis‐loop mediated isothermal amplification assay for rapid and on‐site authentication of buffalo (Bubalus bubalis) meat

Alkaline lysis‐loop mediated isothermal amplification assay for rapid and on‐site authentication of buffalo (Bubalus bubalis) meat

Analytical Methods for Authentication of Buffalo Meat

Rapid Detection of Hepatitis A Virus in Foods Using a Bioluminescent Assay in Real-Time (BART) and Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) Technology

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