Intended use
Clinical information:
The Epstein-Barr virus (EBV; also Human Herpesvirus 4, HHV-4) is a human pathogen, enveloped, double-stranded DNA virus. Infection with the virus happens usually during childhood, without any symptoms. In adolescent or adults it comes to an outbreak of Pfeiffer glandular fever (infectious mononucleosis) in 30-60% of cases. From the 40th about 98% of people are infected with EBV. According to both asymptomatic and symptomatic after infection, the virus persists in the body for life and can be reactivated as all herpes viruses. Usually a reactivation is not noticed by the host and quickly antagonized by the immune system. In case of immunosuppression (eg, HIV or transplant recipients), the virus can multiply uncontrollably and contribute to the emergence of various rare cancers.
RealLine EBV quantitative (Str-format) assay kit is designed to detect Epstein-Barr virus DNA isolated from clinical specimens using extraction kits:
RealLine DNA-Express (REF VBC8899)
RealLine DNA-Extraction 3 (REF VBC8889)
RealLine Extraction 100 (REF VBC8896)
RealLine EBV assay kit is intended for the detection of Epstein-Barr virus DNA in clinical specimens (serum, plasma, cerebrospinal fluid, oropharyngeal swabs, saliva, biopsy materials, bronchopulmonary lavage) and for quantitative determination of EBV DNA in serum (plasma) using the method of real-time polymerase chain reaction (PCR) with fluorescence detection of amplified product.
For a quantitative determination of EBV DNA from blood serum (plasma), use the extraction kits: RealLine DNA-Extraction 2 or RealLine Extraction 100 and follow the instructions below.
The results of PCR analysis are taken into account in complex diagnostics of disease.
The Str-Format Kit contains 96 tubes (0.2 ml) in strips with lyophilized Mastermix. 50 µl of extracted DNA have to be pipetted into the tube and the ready mastermix is diluted. The kit contains reagents required for 96 tests, including control samples and the positive control sample.
The kit is validated for use with iQ™ iCycler, iQ™5 iCycler, CFX96™ (Bio-Rad, USA) DT-96 (DNA-Technology, Russia) and RealLine Cycler (BIORON Diagnostics GmbH).
The use of:
! Extraction Kits for nucleic acids from clinical specimen from other supplier
! other real-time PCR devices
! appropriate reaction volumes, other than 50µl
has to be validated in the lab by the user. The special notes regarding the internal control IC have to be strongly followed.
For optimal results please always use the RealLine Internal Control. In case you do not use RealLine Extraction kits, order RealLine Internal Control (VBC8881) seperately.
Status
CE IvD
Validated extraction kits
RealLine DNA-Express (REF VBC8899)
RealLine DNA-Extraction 3 (REF VBC8889)
RealLine Extraction 100 (REF VBC8896)
Validated real-time PCR platforms
The kit is validated for use with iQ™ iCycler, iQ™5 iCycler, CFX96™ (Bio-Rad, USA) DT-96 (DNA-Technology, Russia) and RealLine Cycler (BIORON Diagnostics GmbH).
Specifications
The range of detectable concentrations (linearity range): 500 to 108 IU EBV DNA/ml.
Specificity of the EBV DNA detection is determined using the Standard Reference Panel of negative sera, SRP NCS, as a percentage of the samples determined by the kit as negative. Specificity equals 100%.
Sensitivity is determined on five samples containing 40 IU of EBV DNA in the sample, prepared from Standard Reference Sample, SRS EBV DNA, as a percentage of the samples, determined by the kit as positive. Sensitivity equals 100%.
The coefficient of variation (CV in %) is calculated for the logarithm values of the EBV DNA concentrations in six SRS samples. The coefficient of variation is not more than 10%.
The “linearity” test characterizes the coincidence (in %) of the measured value and the prescribed value (calculated with allowance for the dilution factor) of the logarithm of the EBV DNA concentration in a sample prepared by dilution from SRS. The “linearity” is in the range of 90-110%.
Diagnostic sensitivity of the EBV DNA detection: clinical tests conducted on 105 positive samples from 80 patients diagnosed with infectious mononucleosis and HIV-infected patients showed 100% sensitivity (interval 96.3% -100%, with a confidence level of 90%).
Diagnostic specificity of the EBV DNA detection: clinical trials conducted on 96 negative samples from nominally healthy blood donors and HIV-infected patients showed 100% specificity (interval 96.9% -100%, with a confidence level of 90%).
Analysis by the CE-marked reference kit showed full match of results.
Order information
| Item number | Kit size | |
| VBD2198 | 96 reactions | |
| RealLine Internal Control | Kit size | |
| VBC8881 | 250 reactions |