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Home / Equine Research Bank / J Clin Microbiol / Fast duplex one-step reverse transcriptase PCR for rapid dif...
Journal of clinical microbiology2010; 48(11); 4010-4014; doi: 10.1128/JCM.00582-10

Fast duplex one-step reverse transcriptase PCR for rapid differential detection of West Nile and Japanese encephalitis viruses.

Abstract: The aim of this study was to develop a highly sensitive and specific one-step duplex reverse transcriptase PCR (RT-PCR) assay for the simultaneous and differential detection of West Nile (WNV) and Japanese encephalitis (JEV) viruses. The bioinformatic analysis of published sequences of WNV and JEV revealed conserved regions not targeted by previously reported primers. A total of 13 primers were designed based on these regions to detect all of the WNV and JEV lineages and to discriminate between the two viruses by the generation of 482- and 241-bp cDNA products, respectively. The results indicate that single-tube duplex PCR using these primers is a useful technique for the detection and differentiation of WNV and JEV in plasma or brain tissue. The novel duplex RT-PCR described in this study enables the early diagnosis of these two encephalitic flaviviruses. In addition, this technique may be useful as part of a testing regimen for human patients, horses, and other susceptible animal species, as it is rapid (less than 3.5 h from RNA extraction), sensitive, and specific, and it may enable the differential diagnosis of clinical samples.
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Publication Date: 2010-09-15 PubMed ID: 20844215PubMed Central: PMC3020880DOI: 10.1128/JCM.00582-10Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research developed a quick and precise testing method for simultaneous identification of West Nile and Japanese encephalitis viruses, allowing for rapid diagnosis in both human and animal samples.

Objective of the Research

  • The primary aim of this research was to develop a sensitive and accurate one-step duplex reverse transcriptase PCR (RT-PCR) testing method. This assay is designed to identify and differentiate between West Nile Virus (WNV) and Japanese Encephalitis Virus (JEV) rapidly.

Methodology and Findings

  • Through their bioinformatics analysis of published West Nile and Japanese Encephalitis viral sequences, the researchers identified conserved regions, which previous primers had not targeted.
  • Based on these conserved regions, the team designed 13 unique primers. They are capable of identifying all known lineages of both the WNV and JEV, and they distinguish between the two viruses using the generation of clear and distinct cDNA products of differing lengths (482-bp for WNV, 241-bp for JEV).
  • The results of this study suggest that this single-tube duplex PCR technique utilizing the newly designed primers is an effective solution for detecting and differentiating WNV and JEV, regardless of whether the sample was drawn from plasma or brain tissue.

Significance and Application of the Research

  • This rapid and effective method of testing enables early detection of these two encephalitis-causing flaviviruses.
  • This duplex RT-PCR method could be instrumental in devising a testing and diagnostic regimen for both human patients and other susceptible animal species such as horses.
  • Due to its speed (taking less than 3.5 hours from RNA extraction), sensitivity, and specificity, physicians and veterinarians could employ this testing method for differential diagnosis of clinical samples, improving patient outcomes by providing faster therapeutic intervention.

Cite This Article

APA
Yeh JY, Lee JH, Seo HJ, Park JY, Moon JS, Cho IS, Lee JB, Park SY, Song CS, Choi IS. (2010). Fast duplex one-step reverse transcriptase PCR for rapid differential detection of West Nile and Japanese encephalitis viruses. J Clin Microbiol, 48(11), 4010-4014. https://doi.org/10.1128/JCM.00582-10

Publication

Journal of clinical microbiology
ISSN: 1098-660X
NlmUniqueID: 7505564
Country: United States
Language: English
Volume: 48
Issue: 11
Pages: 4010-4014

Researcher Affiliations

Yeh, Jung-Yong
  • Foreign Animal Disease Division, National Veterinary Research and Quarantine Service, 480, Anyang-6-Dong, Manan-gu, Anyang-city, Gyeonggi-do 430-824, South Korea. yeh02@nvrqs.go.kr
Lee, Ji-Hye
    Seo, Hyun-Ji
      Park, Jee-Yong
        Moon, Jin-San
          Cho, In-Soo
            Lee, Joong-Bok
              Park, Seung-Yong
                Song, Chang-Seon
                  Choi, In-Soo

                    MeSH Terms

                    • Animals
                    • DNA Primers / genetics
                    • Diagnosis, Differential
                    • Encephalitis Virus, Japanese / genetics
                    • Encephalitis Virus, Japanese / isolation & purification
                    • Encephalitis, Japanese / diagnosis
                    • Encephalitis, Japanese / veterinary
                    • Encephalitis, Japanese / virology
                    • Horses
                    • Humans
                    • Reverse Transcriptase Polymerase Chain Reaction / methods
                    • Sensitivity and Specificity
                    • Virology / methods
                    • West Nile Fever / diagnosis
                    • West Nile Fever / veterinary
                    • West Nile Fever / virology
                    • West Nile virus / genetics
                    • West Nile virus / isolation & purification

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