[Quick test for measurement of rubella virus titer in virus-containing fluid using RT-PCR].
OBJECTIVE
To develop method of rubella virus titer measurement in virus-containing fluid using real-time PCR (RT-PCR) with fluorescent detection.


METHODS
Measurement of infectious titer of rubella virus (Wistar RA 27/3 strain) cultivated on Vero cells was performed simultaneously by RT-PCR and cytopathic effect assay (CEA) on PK-13 cell culture and then results obtained by each method were compared.


RESULTS
Time interval after inoculation, in which difference between virus titer measured by both methods did not exceed 0.3 1gTCD50/ml (value acceptable by WHO), was 2 - 7 days. Pearson correlation coefficient between two values for the mentioned interval was close to 1, which point to good agreement of results. In control sample--international vaccine standard of rubella virus--difference in virus titer determined by RT-PCR and CEA was within 0.2 1gTCD50/ml that lower than value acceptable by WHO.


CONCLUSIONS
Method for measurement of rubella virus titer in virus-containing fluid using RT-PCR was developed, which characterized by high specificity, sensitivity, standard performing, shorter time needed for procedure compared with classic methods and, at the same time, high correlation of its results with results obtained by the latter methods during defined time interval.
Iu I
IuI
Zabiiaka
E B
EB
Faĭzuloev
T K
TK
Borisova
A A
AA
Nikonova
V V
VV
Zverev
D000818:Animals; D002460:Cell Line; D002522:Chlorocebus aethiops; D017931:DNA Primers; D005453:Fluorescence; D016133:Polymerase Chain Reaction / Q000379:methods; D011817:Rabbits; D012409:Rubella / Q000175:diagnosis* / Q000821:virology; D012412:Rubella virus / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D014709:Vero Cells; D019562:Viral Load / Q000379:methods*
D017931:DNA Primers