RealLine CMV quantitative

ASSAY KIT FOR THE QUANTITATIVE DETECTION OF CYTOMEGALOVIRUS (CMV) DNA USING REAL TIME PCR METHOD

 

CE-IvD marked on many real-time PCR devices

 

Ready to use – completely lyophilized

Intended use

RealLine CMV quantitative assay kit is intended for the detection of CMV DNA in clinical specimens (blood serum, plasma, saliva, oropharyngeal swabs, urine, urogenital swabs) using the method  of  real-time  polymerase  chain  reaction  (PCR)  with  fluorescence  detection  of  amplified product.

The kit detects the highly conserved DNA fragment between genes RL1 and RL5A contained in CMV genome in two copies and specific to all known CMV isolates.

The extraction of DNA from clinical material can be performed using the kits:

RealLine DNA-Express (REF VBC8899)

RealLine DNA-Extraction 3 (REF VBC8889)

RealLine Extraction 100 (REF VBC8896)

When using DNA extraction kits of other manufacturers it is highly recommended to use Internal Control sample (IC) manufactured by BIORON Diagnostics GmbH.

For a quantitative determination of CMV DNA from blood serum (plasma), use the extraction kits: 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 (0.2ml) with lyophilized Mastermix. 50µl of extracted DNA have to be pipetted into the tube for rehydration of the lyophilized ready mastermix. The kit contains reagents required for 96 tests, including control samples and the positive control sample.

The kit is validated for use with RealLine Cyclers (BIORON Diagnostics GmbH), iQ™ iCycler, iQ™5 iCycler (Bio-Rad, USA). The kit is compatible with other Real Time PCR systems such as CFX96™ (Bio-Rad, USA) and DT-96 (DNA-Technology, Russia).

The use of:

!  Extraction Kits for nucleic acids from clinical specimen from other supplier

!  other real-time PCR devices

!  appropriate reaction volumes, other than 50 µl

has to be validated in the lab by the user. The special notes regarding the internal control IC have to be strongly followed.

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 RealLine Cyclers (BIORON Diagnostics GmbH), iQ™ iCycler, iQ™5 iCycler (Bio-Rad, USA). The kit is compatible with other Real Time PCR systems such as CFX96™ (Bio-Rad, USA) and DT-96 (DNA-Technology, Russia).

Specifications

The range of detectable concentrations: 300 to 108 IU CMV DNA/ml.

The analytical specificity of RealLine CMV assay kit is ensured by the selection of the primers and probes. The primers and probes were checked for possible homologies to all sequences published in gene banks by sequence comparison analysis.

The  analytical  specificity  of  the  CMV  DNA  detection  is  determined  using  the  Standard Reference  Panel  of  negative  sera  not  containing  causative  agents  of  various  infectious diseases, as a percentage of the samples determined by the kit as negative. Specificity equals 100 %.

The analytical sensitivity is determined on five samples containing 24 IU of CMV DNA in the sample, prepared from Standard Reference Sample, SRS CMV 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 CMV 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  CMV  DNA concentration logarithm in a sample prepared by dilution from SRS. The linearity is in the range of 90-110%

Diagnostic sensitivity of the CMV DNA detection. Clinical  tests  were  conducted  on  153  clinical  samples  from  128  patients  diagnosed  with cytomegalovirus infection (20 samples of whole blood; 47 blood plasma and 25 blood serum samples,  where  25  serum  and  plasma  samples  were  obtained  from  the  same  patients;  8 urogenital swabs, 24 samples of saliva, 16 urine samples).Clinical  tests  showed  100  %  sensitivity  (interval  97.7  %  -100  %,  with  a  confidence  level  of 90 %);

Diagnostic specificity of the CMV DNA detection. Clinical tests were conducted on 92 CMV negative clinical samples from 67 healthy blood donors (20 samples of whole blood; 25 blood plasma and 47 blood serum samples, where 25 serum and plasma samples were obtained from the same patients). Clinical tests showed 100 % specificity (interval 97.4 % -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
VBD1598 96 reactions