Skip to main content
PMC home page
Food Science and Biotechnology logoLink to Food Science and Biotechnology
. 2016 Feb 29;25(1):11–20. doi: 10.1007/s10068-016-0002-1

Nucleic acid-based biotechnologies for food-borne pathogen detection using routine time-intensive culture-based methods and fast molecular diagnostics

Amira Souii 1,2,✉, Manel Ben M’hadheb-Gharbi 2, Jawhar Gharbi 2
PMCID: PMC6049363  PMID: 30263230

Abstract

Diseases caused by food-borne pathogens constitute a major burden to consumers, food business operators, and national governments. Bacterial and viral pathogens are the major biotic factors influencing food safety. A vast array of culture dependent analytical methods and protocols have been developed. Recently, nucleic acid-based methods have begun to replace or complement culture-based methods for routine use in food control laboratories. Basic advantages provided by nucleic acid-based technologies are faster speed and more information, such as sub-species identification, antibiotic resistance, and food microbiology. In particular, PCR and alternative methods have been developed to a stage that provides good speed, sensitivity, specificity, and reproducibility with minimized risk of carryover contamination. This review briefly summarizes currently available and developing molecular technologies that may be candidates for involvement in microbiological molecular diagnostic methods in the next decade.

Keywords: food, diagnostic, pathogen, detection, biotechnology

References

  • 1.Niessen L, Luo J, Denschlag C, Vogel RF. The application of loop-mediated isothermal amplification (LAMP) in food testing for bacterial pathogens and fungal contaminants. Food Microbiol. 2013;36:191–206. doi: 10.1016/j.fm.2013.04.017. [DOI] [PubMed] [Google Scholar]
  • 2.Jacxsens L, Kussaga J, Luning PA, van der Spiegel M, Devlieghere F, Uyttendaele M. A microbial assessment scheme to measure microbial performance of food safety management systems. Int. J. Food Microbiol. 2009;134:113–125. doi: 10.1016/j.ijfoodmicro.2009.02.018. [DOI] [PubMed] [Google Scholar]
  • 3.Newell DG, Koopmans M, Verhoef L, Duizer E, Aidara-Kane A, Sprong H, Opsteegh M, Langelaar M, Threfall J, Scheutz F, van der Giessen J, Kruse H. Food-borne diseases-The challenges of 20 years ago still persist while new ones continue to emerge. Int. J. Food Microbiol. 2010;139:S3–S15. doi: 10.1016/j.ijfoodmicro.2010.01.021. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 4.Gracias KS, McKillip JL. A review of conventional detection and enumeration methods for pathogenic bacteria in food. Can. J. Microbiol. 2004;50:883–890. doi: 10.1139/w04-080. [DOI] [PubMed] [Google Scholar]
  • 5.Hoorfar J. Rapid detection, characterization, and enumeration of food-borne pathogens. APMIS Suppl. 2011;133:1–24. doi: 10.1111/j.1600-0463.2011.02767.x. [DOI] [PubMed] [Google Scholar]
  • 6.Rijpens NP, Herman LMF. Molecular methods for identification and detection of bacterial food pathogens. J. AOAC Int. 2002;85:984–995. [PubMed] [Google Scholar]
  • 7.Scallan E, Hoekstra RM, Angulo FJ, Tauxe RV, Widdowson MA, Roy SL, Jones JL, Griffin PM. Food-borne illness acquired in the United States-Major pathogens. Emerg. Infect. Dis. 2011;17:16–22. doi: 10.3201/eid1701.P21101. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 8.Rodriguez-Lazaro D, Cook N, Ruggeri FM, Sellwood J, Nasser A, Nascimento MSJ, D’Agostino M, Santos R, Saiz JC, Rzezutka A. Virus hazards from food, water and other contaminated environments. FEMS Microbiol. Rev. 2011;36:786–814. doi: 10.1111/j.1574-6976.2011.00306.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 9.Goyal SM. Viruses in Foods. 2006. pp. 1–17. [Google Scholar]
  • 10.Moe CL. Preventing norovirus transmission: How should we handle food handlers? Clin. Infect. Dis. 2008;48:38–40. doi: 10.1086/594119. [DOI] [PubMed] [Google Scholar]
  • 11.Stals A, Baert L, van Coillie E, Uyttendaele M. Extraction of food-borne viruses from food samples: A review. Int. J. Food Microbiol. 2012;153:1–9. doi: 10.1016/j.ijfoodmicro.2011.10.014. [DOI] [PubMed] [Google Scholar]
  • 12.Stals A, van Coillie E, Uyttendaele M. Viral genes everywhere: Public health implications of PCR-based testing of foods. Curr. Opin. Virol. 2013;3:69–73. doi: 10.1016/j.coviro.2012.11.003. [DOI] [PubMed] [Google Scholar]
  • 13.De Medici D, Anniballi F, Wyatt GM, Lindstrom M, Messelhausser U, Aldus CF, Delibato E, Korkeala H, Peck MW, Fenicia L. Multiplex^PCR for detection of botulinum neurotoxin-producing clostridia in clinical, food, and environmental samples. Appl. Environ. Microb. 2009;75:6457–6461. doi: 10.1128/AEM.00805-09. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 14.European Pharmacopoeia. Nucleic Acid Amplification Techniques, 5th ed. Council of Europe, Edqm, Strasbourg, France. pp. 173–176 (2005)
  • 15.Mead PS, Slutsker L, Dietz V, McCaig LF, Bresee JS, Shapiro C, Griffin PM, Tauxe RV. Food-related illness and death in the United States. Emerg. Infect. Dis. 1999;5:607–625. doi: 10.3201/eid0505.990502. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 16.Daniels NA, Bergmire-Sweat DA, Schwab KJ, Hendricks KA, Reddy S, Rowe SM, Fankhauser RL, Monroe SS, Atmar RL, Glass RI, Mead P. A food-borne outbreak of gastroenteritis associated with Norwalk-like viruses: First molecular traceback to deli sandwiches contaminated during preparation. J. Infect. Dis. 2000;181:1467–1470. doi: 10.1086/315365. [DOI] [PubMed] [Google Scholar]
  • 17.Hutin YJ, Pool V, Cramer EH, Nainan OV, Weth J, Williams IT, Goldstein ST, Gensheimer KF, Bell BP, Shapiro CN, Alter MJ, Margolis HS. A multistate, foodborne outbreak of hepatitis A. National Hepatitis A Investigation Team. New Engl. J. Med. 1999;34:595–602. doi: 10.1056/NEJM199902253400802. [DOI] [PubMed] [Google Scholar]
  • 18.Jean J, D’Souza DH, Jaykus LA. Multiplex nucleic acid sequence-based amplification for simultaneous detection of several enteric viruses in model ready-to-eat foods. Appl. Environ. Microb. 2004;70:6603–6610. doi: 10.1128/AEM.70.11.6603-6610.2004. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 19.Rosenfield SI, Jaykus LA. A multiplex reverse transcription polymerase chain reaction method for the detection of food-borne viruses. J. Food Protect. 1999;62:1210–1214. doi: 10.4315/0362-028x-62.10.1210. [DOI] [PubMed] [Google Scholar]
  • 20.Beuret C. Simultaneous detection of enteric viruses by multiplex real-time RTPCR. J. Virol. Method. 2004;115:1–8. doi: 10.1016/j.jviromet.2003.09.005. [DOI] [PubMed] [Google Scholar]
  • 21.Sair AI, D’Souza DH, Jaykus LA. Human enteric viruses as causes of food-borne disease. Compr. Rev. Food Sci. F. 2002;1:73–89. doi: 10.1111/j.1541-4337.2002.tb00008.x. [DOI] [PubMed] [Google Scholar]
  • 22.Gouvea V, Santos N, Carmo-Timenetsky M, Estes MK. Identification of Norwalk virus in artificially seeded shellfish and selected foods. J. Virol. Methods. 1994;48:177–187. doi: 10.1016/0166-0934(94)90117-1. [DOI] [PubMed] [Google Scholar]
  • 23.Leggitt PR, Jaykus LA. Detection methods for human enteric viruses in representative foods. J. Food Protect. 2000;63:1738–1744. doi: 10.4315/0362-028x-63.12.1738. [DOI] [PubMed] [Google Scholar]
  • 24.Compton J. Nucleic acid sequence-based amplification. Natur. 1991;350:91–92. doi: 10.1038/350091a0. [DOI] [PubMed] [Google Scholar]
  • 25.Fusco V, Quero GM. Culture-dependent and culture-independent nucleicacid-based methods used in the microbial safety assessment of milk and dairy products. Compr. Rev. Food Sci. F. 2014;13:493–537. doi: 10.1111/1541-4337.12074. [DOI] [PubMed] [Google Scholar]
  • 26.Yan L, Zhou J, Zheng Y, Gamson AS, Roembke BT, Nakayama S, Sintim HO. Isothermal amplified detection of DNA and RNA. Mol. BioSyst. 2014;10:970–1003. doi: 10.1039/c3mb70304e. [DOI] [PubMed] [Google Scholar]
  • 27.Jean J, Blais B, Darveau A, Fliss I. Detection of hepatitis A virus by the nucleic acid sequence-based amplification (NASBA) technique and comparison with RT-PCR. Appl. Environ. Microb. 2001;67:5593–5600. doi: 10.1128/AEM.67.12.5593-5600.2001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 28.Greene SR, Moe C, Jaykus LA, Cronin M, Grosso L, van Aarle P. Evaluation of the nuclisens basic kit assay for detection of norwalkvirus rna in stool specimens. J. Virol. Method. 2003;108:123–131. doi: 10.1016/S0166-0934(02)00286-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 29.Lunel F, Cresta P, Vitour D, Payan C, Dumont B, Frangeul L, Reboul D, Brault C, Piette JC, Huraux JM. Comparative evaluation of hepatitis C virus RNA quantitation by branched DNA NASBA, and monitor assays. Hepatolog. 1999;29:528–535. doi: 10.1002/hep.510290237. [DOI] [PubMed] [Google Scholar]
  • 30.Chan AB, Fox JD. Nucleic acid sequence-based amplification and other transcription-based amplification methods for research and diagnostic microbiology. Rev. Med. Microbiol. 1999;10:185–196. [Google Scholar]
  • 31.Manojkumar R, Mrudula V. Applications of real-time reverse transcription polymerase chain reaction in clinical virology laboratories for the diagnosis of human diseases. Am. J. Infect. Dis. 2006;2:204–209. doi: 10.3844/ajidsp.2006.204.209. [DOI] [Google Scholar]
  • 32.Fakruddin M, Mazumdar RM, Chowdhury A, Mannan KSB. Nucleic acid sequence based amplification (NASBA)-prospects and applications. Int. J. Life Sci. Pharm. Res. 2012;2:106–121. [Google Scholar]
  • 33.Fakruddin M, Mannan KSB, Chowdhury A, Mazumdar RM, Hossain MN, Islam S, Chowdhury MA. Nucleic acid amplification: Alternative methods of polymerase chain reaction. J. Pharm. Bioall. Sci. 2013;5:245–252. doi: 10.4103/0975-7406.120066. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 34.Jasson V, Jacxsens L, Luning P, Rajkovic A, Uyttendaele M. Alternative microbial methods: An overview and selection criteria. Food Microbiol. 2010;27:710–730. doi: 10.1016/j.fm.2010.04.008. [DOI] [PubMed] [Google Scholar]
  • 35.Rodríguez-Lázaro D, Cook N, Hernández M. Real-time PCR in food science: PCR diagnostics. Curr. Issues Mol. Biol. 2013;15:39–44. [PubMed] [Google Scholar]
  • 36.Kuchta T, Knutsson R, Fiore A, Kudirkiene E, Höhl A, Horvatek Tomic D, Gotcheva V, Pöpping B, Scaramagli S, To Kim A, Wagner M, De Medici D. A decade with nucleic acid-based microbiological methods in safety control of foods. Lett. Appl. Microbiol. 2014;59:263–271. doi: 10.1111/lam.12283. [DOI] [PubMed] [Google Scholar]
  • 37.D’Agostino M, Cook N, Rodriguez-Lazaro D, Rutjes S. Nucleic acid amplification-based methods for detection of enteric viruses: Definition of controls and interpretation of results. Food Environ. Virol. 2011;3:55–60. doi: 10.1007/s12560-011-9063-8. [DOI] [Google Scholar]
  • 38.Stals A, Werbrouck H, Baert L, Botteldoorn N, Herman L, Uyttendaele M, van Coillie E. Laboratory efforts to eliminate contamination problems in the realtime RT-PCR detection of noroviruses. J. Microbiol. Method. 2009;77:72–76. doi: 10.1016/j.mimet.2009.01.018. [DOI] [PubMed] [Google Scholar]
  • 39.Richards GP. Limitations of molecular biological techniques for assessing the virological safety of foods. J. Food Protect. 1999;62:691–697. doi: 10.4315/0362-028x-62.6.691. [DOI] [PubMed] [Google Scholar]
  • 40.Knight A, Li D, Uyttendaele M, Jaykus LA. A critical review of methods for detecting human noroviruses and predicting their infectivity. Crit. Rev. Microbiol. 2013;39:295–309. doi: 10.3109/1040841X.2012.709820. [DOI] [PubMed] [Google Scholar]
  • 41.Fratamico PM, Bagi LK, Cray WC, Narang N, Yan X, Medina M, Liu Y. Detection by multiplex real-time polymerase chain reaction assays and isolation of Shiga toxin-producing Escherichia coli serogroups O26, O45, O103, O111, O121, and O145 in ground beef. Foodborne Pathog. Dis. 2011;8:601–607. doi: 10.1089/fpd.2010.0773. [DOI] [PubMed] [Google Scholar]
  • 42.Mayr AM, Lick S, Bauer J, Tharigen D, Busch U, Huber I. Rapid detection and differentiation of Campylobacter jejuni, Campylobacter coli, and Campylobacter lari in food, using multiplex real-time PCR. J. Food Protect. 2010;73:241–250. doi: 10.4315/0362-028x-73.2.241. [DOI] [PubMed] [Google Scholar]
  • 43.Zhao X, Lin CW, Wang J, Oh DH. Advances in rapid detection methods for food-borne pathogens. J. Microbiol. Biotechnol. 2014;24:297–312. doi: 10.4014/jmb.1310.10013. [DOI] [PubMed] [Google Scholar]
  • 44.Kuchta T, Krascsenicsová K, Bánréti G. Optimization of fluorescence measurement in duplex real-time PCR with taqMan probes labeled with VIC and quenched by TAMRA. Biotechnique. 2007;42:147–149. doi: 10.2144/000112391. [DOI] [PubMed] [Google Scholar]
  • 45.Luz SP, Rodriguez-Valera F, Lan R, Reeves PR. Variation of the ribosomal operon 16S-23S gene spacer region in representatives of Salmonella enterica subspecies. J. Bacteriol. 1998;180:2144–2151. doi: 10.1128/jb.180.8.2144-2151.1998. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 46.Woese CR. Bacterial evolution. Microbiol. Rev. 1987;51:221–271. doi: 10.1128/mr.51.2.221-271.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 47.Woese CR. Prokaryote systematics: The evolution of science. In: Ballows A, editor. The Prokaryotes: A Handbook on the Biology of Bacteria, Ecophysiology. 1992. pp. 3–18. [Google Scholar]
  • 48.Nour M. The 23S ribosomal RNA higher-order structure of leuconostocs. Assoc. Afr. Microbiol. Hyg. Alim. 1999;11:3–12. [Google Scholar]
  • 49.Souii A, Ben Nejma M, Elfray Rhim A, Mastouri M, Bel Hadj Jrad B, Makhlouf M, Nour M. Molecular Identification of 4 Salmonella Serovars isolated from food in Tunisia based on the sequence of the ribosomal RNA genes. Afr. J. Microbiol. Res. 2012;6:6454–6461. doi: 10.5897/AJMR12.964. [DOI] [Google Scholar]
  • 50.Lin CK, Tsen HY. Development and evaluation of two novel oligonucleotide probes based on 16S rRNA sequence for the identification of Salmonella in foods. J. Appl. Bacteriol. 1995;78:507–520. doi: 10.1111/j.1365-2672.1995.tb03093.x. [DOI] [PubMed] [Google Scholar]
  • 51.Pabbaraju K, Miller WL, Sanderson KE. Distribution of intervening sequences in the genes for 23S rRNA and rRNA fragmentation among strains of the Salmonella reference Collection B (SARB) and SARC sets. J. Bacteriol. 2000;182:1923–1929. doi: 10.1128/JB.182.7.1923-1929.2000. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 52.Pabbaraju K, Sanderson KE. Sequence diversity of intervening sequences (IVSs) in the 23S ribosomal RNA in Salmonella spp. Gen. 2000;253:55–66. doi: 10.1016/S0378-1119(00)00239-0. [DOI] [PubMed] [Google Scholar]
  • 53.Tsai CC, Lai CH, Yu B, Tsen HY. Use of specific primers based on the 16S-23S internal transcribed spacer (ITS) region for the screening Bifidobacterium adolescentis in yogurt products and human stool samples. Food Microbiol. 2008;14:219–223. doi: 10.1016/j.ijfoodmicro.2008.08.009. [DOI] [PubMed] [Google Scholar]
  • 54.Magistrali C, Dionisi AM, De Curtis P, Cucco L, Vischi O, Scuota S, Zicavo A, Pezzotti G. Contamination of Salmonella spp. in a pig finishing herd, from the arrival of the animals on the slaughterhouse. Res. Vet. Sci. 2008;85:204–207. doi: 10.1016/j.rvsc.2007.12.002. [DOI] [PubMed] [Google Scholar]
  • 55.Settanni L, Corsetti A. The use of multiplex PCR to detect and differentiate food-and beverage-associated microorganisms: A review. J. Microbiol. Method. 2007;69:1–22. doi: 10.1016/j.mimet.2006.12.008. [DOI] [PubMed] [Google Scholar]
  • 56.Wise MG, Siragusa GR, Plumblee J, Healy M, Cray PJ, Seal BS. Predicting Salmonella enterica serotypes by repetitive sequence-based PCR. J. Microbiol. Method. 2009;76:18–24. doi: 10.1016/j.mimet.2008.09.006. [DOI] [PubMed] [Google Scholar]
  • 57.Jothikumar N, Griffiths MW. Rapid detection of Escherichia coli O157:H7 with multiplex real-time PCR assays. Appl. Environ. Microb. 2002;68:3169–3171. doi: 10.1128/AEM.68.6.3169-3171.2002. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 58.Krascsenicsová K, Piknová L, Kaclíková E, Kuchta T. Detection of Salmonella enterica in food using two-step enrichment and real-time polymerase chain reaction. Lett. Appl. Microbiol. 2008;46:483–487. doi: 10.1111/j.1472-765X.2008.02342.x. [DOI] [PubMed] [Google Scholar]
  • 59.Amagliani G, Omiccioli E, del Campo A, Bruce IJ, Brandi G, Magnani M. Development of a magnetic capture hybridization-PCR assay for Listeria monocytogenes direct detection in milk samples. J. Appl. Microbiol. 2006;100:375–378. doi: 10.1111/j.1365-2672.2005.02761.x. [DOI] [PubMed] [Google Scholar]
  • 60.Chang CC, Chen CC, Wei SC, Lu HH, Liang YH, Lin CW. Diagnostic devices for isothermal nucleic acid amplification. Sensor. 2012;12:8319–8337. doi: 10.3390/s120608319. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 61.Cocolin L, Rantsiou K, Iacumin L, Cantoni C, Comi G. Direct identification in food samples of Listeria spp. and Listeria monocytogenes by molecular methods. Appl. Environ. Microb. 2002;68:6273–6282. doi: 10.1128/AEM.68.12.6273-6282.2002. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 62.Saharan P, Dhingolia S, Khatri P, Duhan JS, Gahlawat SK. Loop mediated isothermal amplification (LAMP) based detection of bacteria: A Review. Afr. J. Biotechnol. 2014;13:1920–1928. doi: 10.5897/AJB2013.13459. [DOI] [Google Scholar]
  • 63.Notomi T, Okayama H, Masubuchi H, Yonekawa T, Watanabe K, Amino N, Hase T. Loop-mediated isothermal amplification of DNA. Nucleic Acids Res. 2000;28:e63. doi: 10.1093/nar/28.12.e63. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 64.Wang F, Jiang L, Yang Q, Prinyawiwatkul W, Ge B. Rapid and specific detection of Escherichia coli serogroups O26, O45, O103, O111, O121, O145, and O157 in ground beef, beef trim, and produce by loop-mediated isothermal amplification. Appl. Environ. Microb. 2012;78:2727–2736. doi: 10.1128/AEM.07975-11. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 65.Shao Y, Zhu S, Jin C, Chen F. Development of multiplex loop-mediated isothermal amplification-RFLP (mLAMP-RFLP) to detect Salmonella spp. and Shigella spp. in milk. Int. J. Food Microbiol. 2011;148:75–79. doi: 10.1016/j.ijfoodmicro.2011.05.004. [DOI] [PubMed] [Google Scholar]
  • 66.Yamazaki W, Kumeda Y, Uemura R, Misawa N. Evaluation of a loop mediated isothermal amplification assay for rapid and simple detection of Vibrio parahaemolyticus in naturally contaminated seafood samples. Food Microbiol. 2011;28:1238–1241. doi: 10.1016/j.fm.2011.04.007. [DOI] [PubMed] [Google Scholar]
  • 67.Nagamine K, Hase T, Notomi T. Accelerated reaction by loop-mediated isothermal amplification using loop primers. Mol. Cell Probe. 2002;16:223–229. doi: 10.1006/mcpr.2002.0415. [DOI] [PubMed] [Google Scholar]
  • 68.Song T, Toma C, Nakasone N, Iwanaga M. Sensitive and rapid detection of Shigella and enteroinvasive Escherichia coli by a loop-mediated isothermal amplification method. FEMS Microbiol. Lett. 2005;243:259–263. doi: 10.1016/j.femsle.2004.12.014. [DOI] [PubMed] [Google Scholar]
  • 69.Fakruddin M, Chowdhury A. Pyrosequencing: An alternative to traditional sanger sequencing. Am. J. Biochem. Biotechnol. 2012;8:14–20. doi: 10.3844/ajbbsp.2012.14.20. [DOI] [Google Scholar]
  • 70.Fakruddin M, Mazumdar RM, Chowdhury A, Hossain MN, Mannan KS. Pyrosequencing-prospects and applications. Int. J. Life Sci. Pharm. Res. 2012;2:65–76. [Google Scholar]
  • 71.Walker GT, Fraiser MS, Schram JL, Little MC, Nadeau JG, Malinowski DP. Strand displacement amplification -An isothermal, in vitro DNA amplification technique. Nucleic Acids Res. 1992;20:1691–1696. doi: 10.1093/nar/20.7.1691. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 72.McHugh TD, Pope CF, Ling CL, Patel S, Billington OJ, Gosling RD, Lipman MC, Gillespie SH. Prospective evaluation of BDProbeTec strand displacement amplification (SDA) system for diagnosis of tuberculosis in non-respiratory and respiratory samples. J. Med. Microbiol. 2004;53:1215–1219. doi: 10.1099/jmm.0.45780-0. [DOI] [PubMed] [Google Scholar]
  • 73.Kolackova I, Karpiskova R. Practical experience with a rapid detection system for Salmonella in food. Seminar, Modern Laboratory Methods in Practice II. 2012. [Google Scholar]
  • 74.Mullis KB. The unusual origin of the polymerase chain reaction. Sci. Am. 1990;262:56–61. doi: 10.1038/scientificamerican0490-56. [DOI] [PubMed] [Google Scholar]
  • 75.Fakruddin M. Loop mediated isothermal amplification-An alternative to polymerase chain reaction (PCR) Bangladesh Res. Pub. J. 2011;5:425–439. [Google Scholar]
  • 76.Wolcott MJ. Advances in nucleic acid-based detection methods. Clin. Microbiol. Rev. 1992;5:370–386. doi: 10.1128/cmr.5.4.370. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 77.Römpler H, Dear PH, Krause J, Meyer M, Rohland N, Schöneberg T, Spriggs H, Stiller M, Hofreiter M. Multiplex amplification of ancient DNA. Nat. Protoc. 2006;1:720–728. doi: 10.1038/nprot.2006.84. [DOI] [PubMed] [Google Scholar]
  • 78.Sakurai A, Shibasaki F. Updated values for molecular diagnosis for highly pathogenic avian influenza virus. Viruse. 2012;4:1235–1257. doi: 10.3390/v4081235. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Food Science and Biotechnology are provided here courtesy of Springer

RESOURCES