241. Development of a Real-Time qPCR Assay for the Detection of Zika Virus RNA: a Surveillance Study
Session: Poster Abstract Session: Diagnostics: Virology
Thursday, October 27, 2016
Room: Poster Hall
  • MM 0499 REV0 1016 2016 ZIKV ID Week.pdf (1.2 MB)
  • Background: Zika virus (ZIKV) was first discovered in 1947 and the first human case confirmed in Uganda in 1952. Before 2015, ZIKV infections were present in tropical Africa, Southeast Asia, and Pacific Islands. The first confirmed Zika virus infection in Brazil was reported in May 2015, and a public health emergency of international concern (PHEIC) was declared by World Health Organization (WHO) on February 1, 2016, attributable to microcephaly in newborns. Increased cases of Guillain-Barré Syndrome post-ZIKV infection have also been observed.

    Methods: We utilized reverse transcription TaqMan® PCR chemistry to detect ZIKV RNA in contrived and clinical specimens. Primers and probes were designed to detect a portion of the envelope gene. Oligonucleotide design achieved specificity and sensitivity in detecting the current as well as other ZIKV strains. This assay utilizes the NucliSens easyMAG Total Nucleic Acid Extraction Platform. RNA is amplified and detected using the TaqPath™ qPCR Master Mix kit on an ABI 7500 SDS Instrument. Analytical performance characteristics were evaluated for each specimen type, and a clinical surveillance study was performed using clinical specimens collected from symptomatic and asymptomatic individuals in Colombia, South America.

    Results: Analytical specificity was confirmed by the absence of signal with closely and distantly related pathogens that present similar signs and symptoms. The limits of detection in plasma and urine were 68 and 211 copies/mL, respectively. Of 153 clinical specimens tested, 23 were positive by qRT-PCR. Seven of these samples were present at sufficiently high titers to allow confirmation of the currently circulating ZIKV strain by Sanger sequencing. A total of 80 specimens from normal U.S. donors were found negative, and 50 samples from the surveillance study returned a negative result.

    Conclusion: This laboratory-developed ZIKV qRT-PCR assay is a sensitive and specific test for detection of ZIKV RNA in plasma, serum, and urine specimens. The availability of this assay expands testing capabilities during the public health emergency, helping to meet public demand for ZIKV testing, which is currently only performed by public health laboratories and a limited number of commercial laboratories.

    James Grantham, BS, Katelyn Bartlett, MS, Ingrid Caton, MS, Steve Kleiboeker, PhD and Michelle Altrich, PhD, Viracor-IBT Laboratories, Lee's Summit, MO


    J. Grantham, Viracor-IBT Laboratories: Employee , Salary

    K. Bartlett, Viracor-IBT Laboratories: Employee , Salary

    I. Caton, Viracor-IBT Laboratories: Employee , Salary

    S. Kleiboeker, Viracor-IBT Laboratories: Employee , Salary

    M. Altrich, Viracor-IBT Laboratories: Employee , Salary

    Findings in the abstracts are embargoed until 12:01 a.m. CDT, Wednesday Oct. 26th with the exception of research findings presented at the IDWeek press conferences.