Novel Molecular Diagnostics and Therapeutic Tools for Livestock Diseases

  • Sushila Maan
  • Sangeeta Dalal
  • Aman Kumar
  • Anita Dalal
  • Nitish Bansal
  • Deepika Chaudhary
  • Akhil Gupta
  • Narender Singh Maan


Recent novelties in diverse diagnostics and therapeutic tools in animal health sector have paved a brighter and clearer way ahead. These are proved to be better in detection, management, control and eradication of animal sufferings caused by various infectious and non-infectious diseases. These innovations have potential impact that extends beyond the animal health and welfare. The advancements have significantly contributed towards improvement in the economy of the country as well as food security. In the present competitive era of evolution, the organisms have inculcated a number of new strategies for survival and spread. Therefore, science needs to continuously evolve more sensitive, specific and high-throughput tools to overcome pathogen cleverness to escape from host immune surveillance.

For visible or remarkable changes, it is necessary to use full potential of these advanced molecular techniques into current animal health standards and practices. Under ‘One Health’ concept, the health of animals and humans has to be taken care simultaneously.

At present, these advanced molecular diagnostic methods play a significant role in the detection of new and emerging pathogens of livestock. The acquired information also helps to study the interrelationships of pathogens, their hosts and their surroundings. Additionally new vaccines bridging human and animal health development may be discovered. Latest developments in the field of diagnostics and vaccine design through genomics approach have also laid the foundation to enhance the diagnosis and surveillance and in turn helped in the control of infectious diseases. Latest high-throughput DNA sequencing platforms are currently being used for identification and detailed analysis of both disease pathogen and host genomes. The high-throughput data generated using these platforms need to be analysed adopting the bioinformatics and computational genomics that have taken a very high pace nowadays. In the context of animal health, the data analysis may provide some key opportunities for the development of better diagnostic and therapeutic tools for emerging or re-emerging diseases. Such novel and potent technologies put forward a significantly new scenario of disease knowledge, which enables more accurate predictions leading to faster and greater management responses to combat potentially devastating disease crises.


  1. Abdullahi UF, Naim R, Aliyu S et al (2015) Loop-mediated isothermal amplification (LAMP), an innovation in gene amplification: bridging the gap in molecular diagnostics; a review. Indian J Sci Technol 8:557–567CrossRefGoogle Scholar
  2. Anderson NL, Anderson NG (1998) Proteome and proteomics: new technologies, new concepts, and new words. Electrophoresis 19:1853–1861CrossRefPubMedGoogle Scholar
  3. Bang D, Church GM (2008) Gene synthesis by circular assembly amplification. Nat Methods 5:37–39CrossRefPubMedGoogle Scholar
  4. Batra K, Kumar A, Kumar V et al (2015) Development and evaluation of loop-mediated isothermal amplification assay for rapid detection of Capripox virus. Vet World 8:1286–1292CrossRefPubMedPubMedCentralGoogle Scholar
  5. Bora M, Bora DP, Barman NN et al (2015) Isolation and molecular characterization of Orf virus from natural outbreaks in goats of Assam. Virus 26:82–88CrossRefGoogle Scholar
  6. Canard B, Sarfati RS (1994) DNA polymerase fluorescent substrates with reversible 3′-tags. Gene 148:1–6CrossRefPubMedGoogle Scholar
  7. Cao Y, Ho DD, Todd J et al (1995) Clinical evaluation of branched DNA signal amplification for quantifying HIV type 1 in human plasma. AIDS Res Hum Retrovir 11:353–361CrossRefPubMedGoogle Scholar
  8. Ceciliani F, Eckersall D, Burchmore R et al (2014a) Proteomics in veterinary medicine: applications and trends in disease pathogenesis and diagnostics. Vet Pathol 51:351–362CrossRefPubMedGoogle Scholar
  9. Ceciliani F, Restelli L, Lecchi C (2014b) Proteomics in farm animals models of human diseases. Proteomics Clin Appl 8:677–688CrossRefPubMedGoogle Scholar
  10. Collins ML, Irvine B, Tyner D et al (1997) A branched DNA signal amplification assay for quantification of nucleic acid targets below 100 molecules/ml. Nucleic Acids Res 25:2979–2984CrossRefPubMedPubMedCentralGoogle Scholar
  11. Compton J (1991) Nucleic acid sequence-based amplification. Nature 350:91–92CrossRefGoogle Scholar
  12. Criado-Fornelio A (2007) A review of nucleic-acid-based diagnostic tests for Babesia and Theileria, with emphasis on bovine piroplasms. Parassitologia 49(Suppl 1):39–44PubMedGoogle Scholar
  13. Datar RH, Joshi NN (2001) Nucleic acids in diagnosis. Natl Med J India 14:34–42PubMedGoogle Scholar
  14. De Leeuw I, Garigliany M, Bertels G et al (2015) Bluetongue virus RNA detection by real-time rt-PCR in post-vaccination samples from cattle. Transbound Emerg Dis 62:157–162CrossRefPubMedGoogle Scholar
  15. Deiman B, van Aarle P, Sillekens P (2002) Characteristics and applications of nucleic acid sequence-based amplification (NASBA). Mol Biotechnol 20:163–179CrossRefPubMedGoogle Scholar
  16. Dilbaghi N, Kaur H, Kumar R et al (2013) Nanoscale device for veterinary technology: trends and future prospective. Adv Mater Lett 4:175–184CrossRefGoogle Scholar
  17. Don RH, Cox PT, Wainwright BJ et al (1991) ‘Touchdown’ PCR to circumvent spurious priming during gene amplification. Nucleic Acids Res 19:4008CrossRefPubMedPubMedCentralGoogle Scholar
  18. Ellington AD, Szostak JW (1990) In vitro selection of RNA molecules that bind specific ligands. Nature 346:818–822CrossRefGoogle Scholar
  19. Erlich HA, Gelfand D, Sninsky JJ (1991) Recent advances in the polymerase chain reaction. Science 252:1643–1651CrossRefPubMedGoogle Scholar
  20. Fakruddin M, Mazumdar RM, Chowdhury A et al (2012) Nucleic acid sequence based amplification (NASBA)-prospects and applications. Int J Life Sci Pharma Res 2:106–121Google Scholar
  21. Gautam R, Mijatovic-Rustempasic S, Esona MD et al (2016) One-step multiplex real-time RT-PCR assay for detecting and genotyping wild-type group A rotavirus strains and vaccine strains (Rotarix(R) and RotaTeq(R)) in stool samples. Peer J 4:e1560CrossRefPubMedGoogle Scholar
  22. Germer K, Leonard M, Zhang X (2013) RNA aptamers and their therapeutic and diagnostic applications. Int J Biochem Mol Biol 4:27–40PubMedPubMedCentralGoogle Scholar
  23. Guatelli JC, Whitfield KM, Kwoh DY et al (1990) Isothermal, in vitro amplification of nucleic acids by a multienzyme reaction modeled after retroviral replication. Proc Natl Acad Sci U S A 87:1874–1878CrossRefPubMedPubMedCentralGoogle Scholar
  24. Johann KS, Schürenkamp M, Sibbing U et al (2015) Linear-after-the-exponential (LATE)-PCR: improved asymmetric PCR for quantitative DNA-analysis. Forensic Sci Int Genet Suppl Ser 5:e659–e661CrossRefGoogle Scholar
  25. Jos A, Pichardo S, Puerto M et al (2009) Cytotoxicity of carboxylic acid functionalized single wall carbon nanotubes on the human intestinal cell line Caco-2. Toxicol in Vitro 23:1491–1496CrossRefPubMedGoogle Scholar
  26. Kellogg DE, Rybalkin I, Chen S et al (1994) TaqStart antibody: “hot start” PCR facilitated by a neutralizing monoclonal antibody directed against Taq DNA polymerase. BioTechniques 16:1134–1137PubMedGoogle Scholar
  27. Kern D, Collins M, Fultz T et al (1996) An enhanced-sensitivity branched-DNA assay for quantification of human immunodeficiency virus type 1 RNA in plasma. J Clin Microbiol 34:3196–3202PubMedPubMedCentralGoogle Scholar
  28. Kim J, Easley CJ (2011) Isothermal DNA amplification in bioanalysis: strategies and applications. Bioanalysis 3:227–239CrossRefPubMedGoogle Scholar
  29. Kwoh DY, Davis GR, Whitfield KM et al (1989) Transcription based amplification system and detection of amplified human immunodeficiency virus type-1 with a bead based sandwich hybridisation format. Proc Natl Acad Sci U S A 86:1173–1177CrossRefPubMedPubMedCentralGoogle Scholar
  30. Lam WY, Yeung AC, Tang JW et al (2007) Rapid multiplex nested PCR for detection of respiratory viruses. J Clin Microbiol 45:3631–3640CrossRefPubMedPubMedCentralGoogle Scholar
  31. Maan NS, Maan S, Belaganahalli M et al (2015) A quantitative real-time reverse transcription PCR (qRT-PCR) assay to detect genome segment 9 of all 26 bluetongue virus serotypes. J Virol Methods 213:118–126CrossRefPubMedGoogle Scholar
  32. Maan S, Maan NS, Batra K et al (2016) Reverse transcription loop-mediated isothermal amplification assays for rapid identification of eastern and western strains of bluetongue virus in India. J Virol Methods 234:65–74CrossRefPubMedGoogle Scholar
  33. Margulies M, Egholm M, Altman WE et al (2005) Genome sequencing in microfabricated high-density picolitre reactors. Nature 437:376–380CrossRefPubMedPubMedCentralGoogle Scholar
  34. Medina C, Santos-Martinez MJ, Radomski A et al (2007) Nanoparticles: pharmacological and toxicological significance. Br J Pharmacol 150:552–558CrossRefPubMedPubMedCentralGoogle Scholar
  35. Mirkin CA, Letsinger RL, Mucic RC et al (1996) A DNA-based method for rationally assembling nanoparticles into macroscopic materials. Nature 382:607–609CrossRefPubMedGoogle Scholar
  36. Mujer CV, Wagner MA, Eschenbrenner M et al (2002) Global analysis of Brucella melitensis proteomes. Ann N Y Acad Sci 969:97–101CrossRefPubMedGoogle Scholar
  37. Mullis KB (1990) The unusual origin of the polymerase chain reaction. Sci Am 262(56–61):64–55Google Scholar
  38. Notomi T, Okayama H, Masubuchi H et al (2000) Loop-mediated isothermal amplification of DNA. Nucleic Acids Res 28:E63CrossRefPubMedPubMedCentralGoogle Scholar
  39. OIE (2016) Manual of standards for diagnostic tests and vaccines. Organization International des Epizootics, Paris, pp 178–188Google Scholar
  40. Pavlov AR, Pavlova NV, Kozyavkin SA, Slesarev AI (2006) Thermostable DNA Polymerases for aWide Spectrum of Applications: comparison of a Robust Hybrid TopoTaq to other enzymes. In: Kieleczawa J (ed) DNA Sequencing II: optimizing preparation and cleanup. Jones and Bartlett, Sudbury, p 241–257. ISBN 0-7637-3383-0Google Scholar
  41. Persing DH (1991) Polymerase chain reaction: trenches to benches. J Clin Microbiol 29:1281–1285PubMedPubMedCentralGoogle Scholar
  42. Pierce KE, Sanchez JA, Rice JE et al (2005) Linear-after-the-exponential (LATE)-PCR: primer design criteria for high yields of specific single-stranded DNA and improved real-time detection. Proc Natl Acad Sci U S A 102:8609–8614CrossRefPubMedPubMedCentralGoogle Scholar
  43. Pratelli A, Decaro N, Tinelli A et al (2004) Two genotypes of canine coronavirus simultaneously detected in the fecal samples of dogs with diarrhea. J Clin Microbiol 42:1797–1799CrossRefPubMedPubMedCentralGoogle Scholar
  44. Raoult D, Aboudharam G, Crubezy E et al (2000) Molecular identification by “suicide PCR” of Yersinia pestis as the agent of medieval black death. Proc Natl Acad Sci U S A 97:12800–12803CrossRefPubMedPubMedCentralGoogle Scholar
  45. Rheem I, Park J, Kim TH et al (2012) Evaluation of a multiplex real-time PCR assay for the detection of respiratory viruses in clinical specimens. Ann Lab Med 32:399–406CrossRefPubMedPubMedCentralGoogle Scholar
  46. Rout MP, Field MC (2001) Isolation and characterization of subnuclear compartments from Trypanosoma brucei. Identification of a major repetitive nuclear lamina component. J Biol Chem 276:38261–38271CrossRefPubMedGoogle Scholar
  47. Ruengwilysup C, Detvisitsakun C, Aumyat N et al (2009) Application of a colony pcr technique with fung’s double tube method for rapid detection and confirmation of clostridium perfringens. J Rapid Method Autom Microbiol 17:280–290CrossRefGoogle Scholar
  48. Sanchez JA, Pierce KE, Rice JE et al (2004) Linear-after-the-exponential (LATE)-PCR: an advanced method of asymmetric PCR and its uses in quantitative real-time analysis. Proc Natl Acad Sci 101:1933–1938CrossRefPubMedGoogle Scholar
  49. Savage N, Thomas T, Duncan J (2007) Nanotechnology applications and implications research supported by the US environmental protection agency star grants program. J Environ Monit 9:1046–1054CrossRefPubMedGoogle Scholar
  50. Schena M, Heller RA, Theriault TP et al (1998) Microarrays: biotechnology's discovery platform for functional genomics. Trends Biotechnol 16:301–306CrossRefPubMedGoogle Scholar
  51. Shah M, Badwaik VD, Dakshinamurthy R (2014) Biological applications of gold nanoparticles. J Nanosci Nanotechnol 14:344–362CrossRefPubMedGoogle Scholar
  52. Shome BR, Das Mitra S, Bhuvana M et al (2011) Multiplex PCR assay for species identification of bovine mastitis pathogens. J Appl Microbiol 111:1349–1356CrossRefPubMedGoogle Scholar
  53. Spargo CA, Dean CH, Nycz CM et al (2000) SDA target amplification. Springer, BerlinCrossRefGoogle Scholar
  54. Torres-Sangiao E, Holban AM, Gestal MC (2016) Advanced nanobiomaterials: vaccines, diagnosis and treatment of infectious diseases. Molecules 21:E867CrossRefPubMedGoogle Scholar
  55. Tsongalis GJ (2006) Branched DNA technology in molecular diagnostics. Am J Clin Pathol 126:448–453CrossRefPubMedGoogle Scholar
  56. Tuerk C, Gold L (1990) Systematic evolution of ligands by exponential enrichment: RNA ligands to bacteriophage T4 DNA polymerase. Science 249:505–510CrossRefGoogle Scholar
  57. Umesha S, Manukumar HM (2016) Advanced molecular diagnostic techniques for detection of food-borne pathogens: current applications and future challenges. Crit Rev Food Sci Nutr 58:1–21Google Scholar
  58. Van Gemen B, Kievits T, Romano J (1995) Transcription based nucleic acid amplification methods like nasba and 3sr applied to viral diagnosis. Rev Med Virol 5:205–211CrossRefGoogle Scholar
  59. Wiedmann M, Wilson WJ, Czajka J et al (1994) Ligase chain reaction (LCR) – overview and applications. PCR Methods Appl 3:S51–S64CrossRefPubMedGoogle Scholar
  60. Wu DY, Wallace RB (1989) The ligation amplification reaction (LAR) – amplification of specific DNA sequences using sequential rounds of template-dependent ligation. Genomics 4:560–569CrossRefPubMedGoogle Scholar
  61. Yamazaki W, Mioulet V, Murray L et al (2013) Development and evaluation of multiplex RT-LAMP assays for rapid and sensitive detection of foot-and-mouth disease virus. J Virol Methods 192:18–24CrossRefPubMedGoogle Scholar
  62. Yatsuda AP, Krijgsveld J, Cornelissen AW et al (2003) Comprehensive analysis of the secreted proteins of the parasite Haemonchus contortus reveals extensive sequence variation and differential immune recognition. J Biol Chem 278:16941–16951CrossRefPubMedGoogle Scholar
  63. Zhan Z, Wei G, Zhu X et al (2009) Development of loop-mediated isothermal amplification (LAMP) method for the detection of peste des petits ruminants virus. J Pathog Biol 4:241–246Google Scholar
  64. Zhang X, Xu S, Gao X et al (2008) The application of asymmetric PCR-SSCP in gene mutation detecting. Front Agric China 2:361–364CrossRefGoogle Scholar
  65. Zhou J, Tompson DK (2004) Microarray technology and applications in environmental microbiology. Adv Agron 82:183–270CrossRefGoogle Scholar

Copyright information

© Springer Nature Singapore Pte Ltd. 2018

Authors and Affiliations

  • Sushila Maan
    • 1
  • Sangeeta Dalal
    • 2
  • Aman Kumar
    • 1
  • Anita Dalal
    • 3
  • Nitish Bansal
    • 1
  • Deepika Chaudhary
    • 1
  • Akhil Gupta
    • 1
    • 3
  • Narender Singh Maan
    • 4
  1. 1.Department of Animal Biotechnology, College of Veterinary SciencesLala Lajpat Rai University of Veterinary and Animal Sciences (LUVAS)HisarIndia
  2. 2.State Cattle Breeding ProjectHisarIndia
  3. 3.Department Veterinary Microbiology, College of Veterinary SciencesLala Lajpat Rai University of Veterinary and Animal Sciences (LUVAS)HisarIndia
  4. 4.Resource Faculty, Department of Animal Biotechnology, College of Veterinary SciencesLala Lajpat Rai University of Veterinary and Animal Sciences (LUVAS)HisarIndia

Personalised recommendations