Skip to main content

Advertisement

SpringerLink
Log in

Enzyme-free amplification and detection of bovine viral diarrhea virus RNA using hybridization chain reaction and gold nanoparticles

  • Methods and protocols
  • Published:
Applied Microbiology and Biotechnology Aims and scope Submit manuscript

Abstract

A novel bovine viral diarrheal virus (BVDV)-RNA detection method was developed using a combination of the amplification capability of hybridization chain reaction (HCR) with the sensitivity of an unmodified-gold nanoparticle (AuNP) colorimetric detection assay. Two auxiliary probes were designed to target a conserved RNA sequence among the BVDV isolates. The complementary target BVDV-RNA was used as the initiator to trigger a cascade of hybridization events to yield nicked double-helix DNA analogous to the alternating copolymers. DNA in the form of a nicked double helix did not prevent salt-induced aggregation of AuNPs. In contrast, in the absence of the complementary target BVDV-RNA, free hairpins with single-stranded sticky ends adsorbed onto the AuNPs, stabilize them, and prevent salt-induced aggregation of the AuNP. The limit of detection (LOD) for the BVDV-RNA was estimated to be 0.008 tissue culture infective dose (TCID50)/reaction. The method developed was highly selective and specific to detect BVDV isolates in clinical samples. This protocol offers a rapid, simple, and cost-effective assay for detecting BVDV.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Scheme 1
Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  • Aebischer A, Beer M, Hoffmann B (2014a) Development and validation of rapid magnetic particle based extraction protocols. Virol J 11:1–9

    Article  Google Scholar 

  • Aebischer A, Wernike K, Hoffmann B, Beer M (2014b) Rapid genome detection of Schmallenberg virus and bovine viral diarrhea virus by use of isothermal amplification methods and high-speed real-time reverse transcriptase PCR. J Clin Microbiol 52:1883–1892

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bachofen C, Bollinger B, Peterhans E, Stalder H, Schweizer M (2013) Diagnostic gap in Bovine viral diarrhea virus serology during the periparturient period in cattle. J Vet Diagn Investig 25:655–661

    Article  Google Scholar 

  • Bi S, Zhao T, Luo B, Zhu JJ (2013) Hybridization chain reaction-based branched rolling circle amplification for chemiluminescence detection of DNA methylation. Chem Commun 49:6906–6908

    Article  CAS  Google Scholar 

  • Chemeris DA, Nikonorov YM, Vakhitov VA (2008) Real-time hybridization chain reaction. Dokl Biochem Biophys 419:53–55

    Article  CAS  PubMed  Google Scholar 

  • Chen Y, Xu J, Su J, Xiang Y, Yuan R, Chai Y (2012) In situ hybridization chain reaction amplification for universal and highly sensitive electrochemiluminescent detection of DNA. Anal Chem 84:7750–7755

    Article  CAS  PubMed  Google Scholar 

  • Choi HMT, Beck VA, Pierce NA (2014) Next-generationin situ hybridization chain reaction: higher gain, lower cost, greater durability. ACS Nano 8:4284–4294

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Dirks RM, Pierce NA (2004) Triggered amplification by hybridization chain reaction. Proc Natl Acad Sci U S A 101:15275–15278

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Dong J, Cui X, Deng Y, Tang Z (2012) Amplified detection of nucleic acid by G-quadruplex based hybridization chain reaction. Biosens Bioelectron 38:258–263

    Article  CAS  PubMed  Google Scholar 

  • Dubovi EJ (2013) Laboratory diagnosis of bovine viral diarrhea virus. Biologicals 41:8–13

    Article  PubMed  Google Scholar 

  • Gilbert SA, Burton KM, Prins SE, Deregt D (1999) Typing of bovine viral diarrhea viruses directly from blood of persistently infected cattle by multiplex PCR. J Clin Microbiol 37:2020–2023

    CAS  PubMed  PubMed Central  Google Scholar 

  • Guo Q, Chen Y, Song Z, Guo L, Fu F, Chen G (2014) Label-free and enzyme-free sensitive fluorescent detection of human immunodeficiency virus deoxyribonucleic acid based on hybridization chain reaction. Anal Chim Acta 852:244–249

    Article  CAS  PubMed  Google Scholar 

  • Ho HA, Najari A, Leclerc M (2008) Optical detection of DNA and proteins with cationic polythiophenes. Acc Chem Res 41:168–178

    Article  CAS  PubMed  Google Scholar 

  • Huang J, Wu Y, Chen Y, Zhu Z, Yang X, Yang CJ, Wang K, Tan W (2011) Pyrene-excimer probes based on the hybridization chain reaction for the detection of nucleic acids in complex biological fluids. Angew Chem Int Ed Engl 50:401–404

    Article  CAS  PubMed  Google Scholar 

  • Jiang Y, Li B, Chen X, Ellington AD (2012) Coupling two different nucleic acid circuits in an enzyme-free amplifier. Molecules 17:13211–13220

    Article  CAS  PubMed  Google Scholar 

  • Li H, Rothberg L (2004) Colorimetric detection of DNA sequences based on electrostatic interactions with unmodified gold nanoparticles. PNAS 101:14036–14039

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Li B, Ellington AD, Chen X (2011) Rational, modular adaptation of enzyme-free DNA circuits to multiple detection methods. Nucleic Acids Res 39:1–13

    Article  Google Scholar 

  • Liu P, Yang X, Sun S, Wang Q, Wang K, Huang J, Liu J, He L (2013) Enzyme-free colorimetric detection of DNA by using gold nanoparticles and hybridization chain reaction amplification. Anal Chem 85:7689–7695

    Article  CAS  PubMed  Google Scholar 

  • Luzzago C, Lauzi S, Ebranati E, Giammarioli M, Moreno A, Cannella V, Masoero L, Canelli E, Guercio A, Caruso C, Ciccozzi M, De Mia GM, Acutis PL, Zehender G, Peletto S (2014) Extended genetic diversity of bovine viral diarrhea virus and frequency of genotypes and subtypes in cattle in Italy between 1995 and 2013. Biomed Res Int 2014:147145

    Article  PubMed  PubMed Central  Google Scholar 

  • Ma C, Wang W, Li Z, Cao L, Wang Q (2012) Simple colorimetric DNA detection based on hairpin assembly reaction and target-catalytic circuits for signal amplification. Anal Biochem 429:99–102

    Article  CAS  PubMed  Google Scholar 

  • Mirkin CA, Letsinger RL, Mucic RC, Storhoff JJ (1996) A DNA-based method for rationally assembling nanoparticles into macroscopic materials. Nature 382:607–609

    Article  CAS  PubMed  Google Scholar 

  • Niu S, Jiang Y, Zhang S (2010) Fluorescence detection for DNA using hybridization chain reaction with enzyme-amplification. Chem Commun 46:3089–3091

    Article  CAS  Google Scholar 

  • Shirvani E, Lotfi M, Kamalzadeh M, Noaman V, Bahriari M, Morovati H, Hatami A (2012) Seroepidemiological study of bovine respiratory viruses (BRSV, BoHV-1, PI-3V, BVDV, and BAV-3) in dairy cattle in central region of Iran (Esfahan province). Trop Anim Health Prod 44:191–195

    Article  PubMed  Google Scholar 

  • Song C, Yang X, Wang K, Wang Q, Huang J, Liu J, Liu W, Liu P (2014) Label-free and non-enzymatic detection of DNA based on hybridization chain reaction amplification and dsDNA-templated copper nanoparticles. Anal Chim Acta 827:74–79

    Article  CAS  PubMed  Google Scholar 

  • Spiga FM, Bonyár A, Ring B, Onofri M, Vinelli A, Sántha H, Guiducci C, Zuccheri G (2014) Hybridization chain reaction performed on a metal surface as a means of signal amplification in SPR and electrochemical biosensors. Biosens Bioelectron 54:102–108

    Article  CAS  PubMed  Google Scholar 

  • Thaxton CS, Georganopoulou DG, Mirkin CA (2006) Gold nanoparticle probes for the detection of nucleic acid targets. Clin Chim Acta 363:120–126

    Article  CAS  PubMed  Google Scholar 

  • Zhang B, Liu B, Tang D, Niessner R, Chen G, Knopp D (2012) DNA-based hybridization chain reaction for amplified bioelectronic signal and ultrasensitive detection of proteins. Anal Chem 84:5392–5399

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

Financial support of this study was provided by Shahid Chamran University of Ahvaz.

Author information

Authors and Affiliations

  1. Department of Biology, Faculty of Science, Shahid Chamran University of Ahvaz, Ahvaz, 6135743135, Iran

    Shahrzad Ghasemi Monjezi & Seyedeh Elham Rezatofighi

  2. Department of animal science and food, Khouzastan Ramin Agriculture and Natural Resources University, Ahvaz, Iran

    Khalil Mirzadeh

  3. Department of Chemistry, Faculty of Science, Shahid Chamran University of Ahvaz, Ahvaz, Iran

    Saadat Rastegarzadeh

Authors
  1. Shahrzad Ghasemi Monjezi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Seyedeh Elham Rezatofighi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Khalil Mirzadeh
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Saadat Rastegarzadeh
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Seyedeh Elham Rezatofighi.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Compliance with ethics requirements

This article does not contain any studies with human or animal subjects.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Ghasemi Monjezi, S., Rezatofighi, S.E., Mirzadeh, K. et al. Enzyme-free amplification and detection of bovine viral diarrhea virus RNA using hybridization chain reaction and gold nanoparticles. Appl Microbiol Biotechnol 100, 8913–8921 (2016). https://doi.org/10.1007/s00253-016-7785-0

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00253-016-7785-0

Keywords

Search

Navigation