Zika detection: comparison of methodologies

Abstract

Many countries in the Americas have detected local transmission of multiple arboviruses that cause febrile illnesses. Therefore, laboratory testing has become an important tool for confirming the etiology of these diseases. The present study aimed to compare the sensitivity and specificity of three different Zika virus detection assays. One hundred serum samples from patients presenting with acute febrile symptoms were tested using a previously reported TaqMan® RT-qPCR assay. We used a SYBR® Green RT-qPCR and a conventional PCR methodologies to compare the results. Of the samples that were determined to be negative by the TaqMan® RT-qPCR assay, 100% (Kappa = 0.670) were also found to be negative by SYBR® Green RT-qPCR based on Tm comparison; however, 14% (Kappa = 0.035) were found to be positive by conventional PCR followed by agarose gel electrophoresis. The differences between the ZIKV strains circulating worldwide and the low viremia period can compromise diagnostic accuracy and thereby the accuracy of outbreak data. Therefore, improved assays are required to improve the diagnosis and surveillance of arbovirus.

Zika virus serological diagnosis: commercial tests and monoclonal antibodies as tools

“…Diagnostic tests can consist of molecular or serological assays. The former are direct assays used for detection and/or quantification of genetic variants, i.e., they are based on the presence of viral nucleic acids in bodily fluids [ 30 ]. In contrast, serological assays can be employed to indirectly identify the previous circulating virus and to measure the patient's immune response against the virus by detecting antibodies against the virus in serum.…”

Phylogenetic surveillance of travel-related Zika virus infections through whole-genome sequencing methods

“…Surveillance of ZIKV is essential to the development of public health guidelines to prevent recurring and emerging ZIKV outbreaks, which are associated with growing numbers of neurological abnormalities and birth malformations 20,21 . Currently, ZIKV is detected through serological testing and real time RT-PCR, and these methods may not always be reliable 22,23 . In this proof-of-concept study, we performed WGS on five Canadian samples from the BCCDC with confirmed travel-acquired ZIKV infections, and aimed to verify the original countries of infections.…”

Descripción de pruebas moleculares en el diagnóstico del virus Zika en el periodo 2008-Febrero 2018. Revisión sistemática

“…Con relación a las pruebas moleculares para el diagnóstico de ZIKV que se han publicado hasta la fecha, se encontró que las pruebas más frecuentemente utilizadas son la RT-PCR convencional y la RT-PCR en tiempo real para amplificar 9 blancos moleculares diferentes. Ambas metodologías son las más recomendadas para amplificar virus tipo RNA como es el caso del ZIKV y son pruebas que presentan una alta sensibilidad (94-96%) y especificidad (97-100%) (34).…”

“…Por otra parte, al identificar los componentes moleculares que permiten la detección del ZIKV, aun cuando en cada una de las técnicas utilizadas hay una gran variabilidad en cuanto al blanco molecular de interés, se determinó que los principales blancos moleculares son los genes E, Ns1 y Ns5, siendo el gen E el de mayor predominio y de uso común en todas las técnicas ya que es el gen encargado de la adherencia y fusión con la membrana. (Ver Tabla1) (15,34,50).…”