Detection of polyomavirus JC genotype from transplant patients by capillary electrophoresis: comparison to fragment length polymorphism analysis.
OBJECTIVE
Among the genotypes of human polyomavirus JC (JCV) reported in Taiwan, CY, TW1, TW2 and TW3 are the most commonly correlated with human diseases. JCV is usually detected using nucleotide sequencing and restriction fragment length polymorphism (RFLP) analysis. The aim of this study was to detect the rate of positivity and genotype of the JCV genome in urine by RFLP or capillary electrophoresis (CE) in renal transplant patients and healthy volunteers.


METHODS
We compared CE analysis to the methods of nucleotide sequencing and RFLP analysis for detection of JCV viruria among 60 renal transplant patients and 50 unrelated healthy controls. Genotyping of the positive PCR products was performed using CE and RFLP analysis simultaneously.


RESULTS
The urine JCV-positive rate was significantly higher in renal transplant patients than in healthy volunteers (40% [24/60] vs. 20% [10/50]; p=0.0238). In addition, multiple genotypes of JCV could be detected by CE, but only one genotype could be detected by RFLP. In our study, 20% (2/10) of urine JCV-positive samples from healthy volunteers had two different genotypes. In renal transplant patients 66% (16/24) of JCV-positive samples had two different genotypes and 12% (3/24) had three different genotypes.


CONCLUSIONS
In comparison with RFLP, CE can detect multiple genotypes in urine JCV-positive samples and requires only 1/200 of the volume of specimen required for RFLP analysis. The CE method has sensitivity and specificity suitable for use in the clinical laboratory, and identifies more genotypes than RFLP analysis.
Lai
Ning-Sheng
NS
Department of Rheumatology, Buddhist Dalin Tzu Chi General Hospital, Chia-Yi, Taiwan.
Lu
Ming-Chi
MC
Lee
Ming-Che
MC
Lin
Teng-Yi
TY
Yin
Wen-Yao
WY
D000328:Adult; D019075:Electrophoresis, Capillary / Q000379:methods*; D005838:Genotype; D006801:Humans; D007577:JC Virus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D016030:Kidney Transplantation; D012150:Polymorphism, Restriction Fragment Length; D012680:Sensitivity and Specificity