Identification of pig DNA using loop-mediated isothermal amplification

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Published: Jun 30, 2017
Keywords:
LAMP pig DNA identification mitochondrial DNA cytochrome b gene

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Nattakorn Rungruang
Faculty of Science and Technology, Rajamangala University of Technology Thanyaburi
Pongsathon Phapugrangkul
Thailand Institute of Scientific and Technological Research
Dolnapa Kaewpa
Faculty of Science and Technology, Rajamangala University of Technology Thanyaburi

Abstract

Loop-mediated isothermal amplification (LAMP) assay and LAMP primers were developed in this study to detect mitochondrial DNA (mtDNA) of pig. The specific LAMP primers were designed from cytochrome b (cyt b) mtDNA of pig that composed of two inner primers (FIP, BIP) and outer primers (F3, B3). The FIP and BIP primers are tandem primers used for loop amplification. The FIP primer set is composed of the forward primer (F2) and the reverse F1c primer linked by additional 4 nucleotide bases, TTTT. The BIP primer set is composed of the reverse primer (B2) and the forward B1c primer linked by TTTT. All LAMP primers recognized six separate regions within target DNA. The cyt b mtDNA was amplified at a range of temperature 57-60oC using Bst DNA polymerase for 60 min with 1 ng of DNA from meat samples. The specificity of LAMP primers was performed using DNA from four meat species (pork, cattle, chicken and duck). The LAMP product was successfully amplified only from cyt b gene of pig whereas no amplified genes was detected from other species. These suggest that the LAMP assay studied was suitable for identifying pig DNA in meat samples.

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How to Cite
1.
Rungruang N, Phapugrangkul P, Kaewpa D. Identification of pig DNA using loop-mediated isothermal amplification. Prog Appl Sci Tech. [Internet]. 2017 Jun. 30 [cited 2024 Apr. 30];7(1):34-42. Available from: https://ph02.tci-thaijo.org/index.php/past/article/view/243109
Issue
Vol. 7 No. 1 (2017): January - June 2017
Section
Biology and Bioresource technology

References

Murugaiah C., Noor Z.M., Mastakim M., Bilung L.M., Selamat J. and Radu S. Meat species identification and halal authentication analysis using mitochondrial DNA. Meat Sci. 2009. 83(1): 57-61.

Khadijah N., Yaakob B. C. M. and Awis Q. S. Hala authenticity issue in meat and meat products. Meat Sci. 2012. 91: 207-214.

Dooley J. J., Paine K. E., Garrett S. D. and Brown H. M. Detection of meat species using TaqMan real-time PCR assay. Meat Sci. 2004. 68:431-438.

Erwanto Y., Abidin M.Z., Sismindari and Rohman A. Pig species identification in meatballs using polymerase chain reaction-restriction fragment length polymorphysm for Halal authentication. Int Food Res J. 2012. 19(3): 901-906.

Avise J. C., Arnol J., Ball R. M., Bermingham E., Lamb T., Neigel J. E., Reeb C. A. and Saunders N. C. Intraspecific phylogeography-the mitrochondrial-DNA bridge between population-genetics and systematics. Annu Rev. Ecol. Syst. 1987. 18: 489-522.

Koh B. R. D., Kim J. Y., Na H. M., Park S. D. and Kim Y. H. Development of species-specific multiplex PCR assays of mitochondrial 12S rRNA and 16S rRNA for the identification of animal species. Korean J. Vet. Serv. 2011. 34:417-428.

Moti Y. and Notomi T. Loop-mediated isothermal amplification (LAMP): a rapid, accurate, and cost-effective diagnostic method for infectious diseases. J Infect Chemother. 2009. 15: 62-69.

Notomi T., Okayama H., Masubichi H., Yonekawa T., Watanabe k., Amino N., Hase T. Loop-mediated isothermal amplification of DNA. Nucleic acid Res. 2000. 28(12): E63.

Enosawa M., Kageyama S., Sawai K., Watanabe K., Notomi T., Onoe S., Mori Y., Yokomizo Y. Use of loop-mediated isothermal amplification of the IS900 sequence fo rapid detection of cultured Mycobacreium avium subsp. paratuberculosis. J Clin Microbiol. 2003. 41(9): 4359-4365.

Misawa Y., Yoshida A., Saito R., Yoshida H., Okuzumi K., Ito N., Okada M., Moriya K., Koike K. Application of loop-mediated isothermal amplification technique to rapid and direct detection of methicillin-resistant Staphylococcus aureus (MRSA) in blood culture. J Infect Chemother. 2007. 13(3): 134-140.

Ohtsuka K., Yanagawa K., Takatori K, Hara-Kudo Y. Detection of Salmonella enterica in naturally contaminated liquid eggs by loop-mediated isothermal amplification, and characterization of Salmonella isolates. Appl Environ Microbiol. 2005. 71(11): 6730-6735.

Cai T., Lou G., Yang J., Xu D., Meng Z. Development and evaluation of real-time loop-mediated isothermal amplification for hepatitis B virus DNA quantification: a new tool for HBV management. J Clin Virol. 2008. 41(4): 2700-276.

Imai M., Ninomiya A., Minekawa H., Notomi T., Ishizaki T., Van Tu P., Tien NT., Tashiro M., Odagiri T. Rapid diagnosis of H5N1 avian influenza virus infection by newly developed influenza H5 hemagglutinin method. J Virol Methods. 2007. 141(2): 173-180.

Poon LL., Leung CS., Chan KH., Lee JH., Yuen KY., Guan Y., Peiris JS. Detection of human influenza A viruses by loop-mediated isothermal amplification method. J Clin Microbiol. 2005. 43(1): 427-430.

Wang YL., Yuan XY., Li TF., Yu KX., Xu HY., Zhang YX., Yu KZ., Liao M., Qin ZM. Rapid detection of newly isolated Tembusu-related Flavivirus by reverse-transcription loop-mediated isothermal amplification assay. Virol J. 2011. 8:533. doi: 10.1186/1743-422X-8-553.

Yoneyama T., Kiyohara T., Shimasaki N., Kobayashi G., Ota Y., Notomi T., Totsuka A., Wakita T. Rapid and real-time detection of hapatitis A virus by reverse transcription loop-mediated isothermal amplification assay. J Virol Methods. 2007. 145(2): 162-168.

Chen R., Tong Q., Zhang Y., Lou D., Kong Q, Lv S., Zhuo M., Wen L., Lu S. Loop-mediated isothermal amplification: rapid detection of Angiostrongyrus cantonensis infection in Pomacea canaliculata. Parasites Vectors. 2011. 4: 204.

Han ET., Watanabe R., Sattabongkot J., Khuntirat B., Sirichaisinthop J., Iriko H., Jin L., Takeo S., Tsuboi T. Detection of four Plasmodium species by genus- and species-specific loop-mediated isothermal amplification for clinical diagnosis. J Clin Microbiol. 2007. 45(8) 2521-2528.

Kong QM., Lu SH., Tong QB., Lou D., Chen R., Zheng B., Kumagai T., Wen LY., Ohta N., Zhou XN. Loop-mediated isothermal amplification (LAMP): early detection of Toxoplasma gondii infection in mice. Parasites Vectors. 2012;5:2. doi:10.1186/1756-3305-5-2.

Guan XY., Guo JC., Shen P., Yang LT., Zhang DB. Visual and rapid detection of two genetically modified soy bean events using loop-mediated isothermal amplification method. Food Anal Methods. 2010. 3(4): 313-320.

Kiddle G., Hardinge P., Buttigieg N., Gandelman O., Pereira C., McElgunn CJ., Rizzoli M., Jackson R., Appleton N., Moore C., Tisi LC., Murray JA. GMO detection using a bioluminescent real time reporter (BART) of loop-mediated isothermal amplification (LAMP) suitable for field use. BMC Biotechnol. 2012. 12:15. doi:10.1186/1472-6750-12-15.

Lee D., La Mura M., Allnutt TR., Powell W. Detection of genetically modified organisms (GMO) using isothermal of target DNA sequences. BMC Biotechnol. 2009. 9:7. doi:10.1186/1472-6750-9-7.

Liu M., Luo Y., Tao R., He R., Jiang K., Wang B., Wang L. Sensitive and rapid detection of genetically modified soybean (Roundup Ready) by loop-mediated isothermal amplification. Biosci Biotechnol Biochem. 2009. 73(11): 2365-2369.

Tindall KR. and Kunkel TA. Fidelity of DNA synthesis by the Thermus aquaticus DNA polymerase. Biochemistry. 1988. 27(16): 6008-6013.