Because of the high specificity and sensitivity, reverse transcriptase polymerase chain reaction (RT-PCR
) was widely utilized in laboratories.
This new RT-PCR
method will help in general detection in first PCR reaction (products of this PCR can be used for direct sequencing) while typing in second multiplex PCR.
Ten-fold serial dilutions of ZikaSPH2015 strain (7) ranging from 106-100 copies/mL, titrated in Vero cells, were spiked in a serum sample from a healthy donor and were tested by real-time RT-PCR
We detected Zika virus by real time RT-PCR
in both the transport medium in which the chorionic biopsy was stored ([C.sub.t] = 36) and the supernatant of the karyotype cell culture ([C.sub.t] = 12).
All 1296 samples, include 34 CSF and 1262 plasma/ serum specimen, were tested over a period of 4 months (September-December 2016) for the presence of CHIK virus-specific RNA by RT-PCR
. Positive and negative controls were included in each run of the assays, and all precautions to prevent cross-contamination were observed.
During 2016, tissue specimens from 627 completed pregnancies with possible maternal Zika virus exposure from the 50 states and DC were submitted to CDC and were tested by Zika virus tissue RT-PCR
. These specimens included placental tissues from 546 live births and placental and/or fetal tissues from 81 pregnancy losses; IHC testing for Zika virus was also performed on specimens from 91 live births and pregnancy losses (15%), criteria for which are specified below.
Up to 2012 two RT-PCR
studies were found, both published in 2011, one of which was published by Kalantri et al.
Three qRT-PCRs were evaluated in this study, RealStar Dengue RT-PCR
, GenoAmp Trioplex Real-Time RT-PCR
Zika/Den/Chiku, and GenoAmp Real-Time RT-PCR
During the development of this protocol, assessments and comparisons of alternative experiment designs were also conducted for different HIV viral RNA extraction approaches (manual extraction with QIAamp Viral RNA extraction kit versus silica bead-based HIV RNA extraction using NucliSens easyMAG system (BioMerieux, Canada)), different RT-PCR
protocols (one-step versus two-step RTPCR), and different nested PCR protocols (one-amplicon approach as described above versus two-amplicon approach which amplifies the HIV-1 PR and RT gene separately) along with various volumes and concentrations of the reagents, templates, or primers used.
Of the 24 women with positive RT-PCR
results and adverse pregnancy outcomes, 23 had symptom onset during the first trimester, while all 8 patients with positive RT-PCR
results but healthy infants had symptom onset in the third trimester (P less than .0001).
In a study by Wang et al in 2010 dengue infection was detected by either MAC-ELISA or RT-PCR
in 300 samples (93.75%), by MAC-ELISA in 229 samples (71.56%), and by RT-PCR
in 162 samples (50.63%)8.
In conjunction, the company performs testing for its proprietary Zika RT-PCR
test as well as the CDC-based Zika MAC-ELISA test at its Focus Diagnostics laboratory in San Juan Capistrano, California as well as performs the molecular RT-PCR
Zika test for use on serum and paired urine specimens at its laboratory in Chantilly, Virginia.