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Loop-Mediated Isothermal Amplification (LAMP) for Detection of Ureaplasma diversum from Cervico-vaginal Swab of Buffaloes

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Abstract

The main plan of the current study was to develop a rapid, robust, and field-applicable loop-mediated isothermal amplification (LAMP) assay for the detection of Ureaplasma diversum. A strain-specific 16S rRNA gene of Ureaplasma diversum was used for detection which was cloned, sequenced, and characterized earlier. LAMP results were visualized within 90 min with the naked eye. Cervico-vaginal swabs of 50 buffaloes were randomly collected from Livestock Research Center of NDRI as per the Institute Animal ethics guidelines. Out of 50 cervico-vaginal swab samples collected randomly, 34 were found positive with LAMP while 16 samples were negative. Conventional PCR results showed the same result. Therefore, the accuracy of the developed LAMP was about 100%. The developed LAMP assay can also be used to screen the animals for Ureaplasma diversum infection in cervico-vaginal swab. However, further study is needed to assess sensitivity and accuracy towards their detection and their relationship in disease diagnosis.

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Acknowledgments

The authors acknowledge Dr. Subhash Chand, T.O at LRC, NDRI, for his support in the sample collection.

Funding

This work was supported by the Department of Biotechnology, Ministry of Science and Technology, New Delhi, India.

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Authors and Affiliations

  1. Animal Biochemistry Division, National Dairy Research Institute, Karnal, Haryana, 132001, India

    Sanjay Sharma, Mamta Pandey, Suneel Kumar Onteru & Dheer Singh

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  1. Sanjay Sharma
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  2. Mamta Pandey
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  3. Suneel Kumar Onteru
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  4. Dheer Singh
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Corresponding author

Correspondence to Dheer Singh.

Ethics declarations

All the applicable institutional guidelines for the care and use of animals were followed. Permission for the collection of the cervico-vaginal swabs used in the present study was approved by Institutional Animal Ethics Committee, NDRI, Karnal, and Haryana, India (Vide no. 41-IAEC-18-1).

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Sharma, S., Pandey, M., Onteru, S.K. et al. Loop-Mediated Isothermal Amplification (LAMP) for Detection of Ureaplasma diversum from Cervico-vaginal Swab of Buffaloes. Appl Biochem Biotechnol 190, 1201–1211 (2020). https://doi.org/10.1007/s12010-019-03155-2

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  • DOI: https://doi.org/10.1007/s12010-019-03155-2

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