Detection of BK virus and JC virus DNA in urine samples from immunocompromised (HIV-infected) and immunocompetent (HIV-non-infected) patients using polymerase chain reaction and microplate hybridisation.
BACKGROUND
The majority of the human population is infected with two human polyomaviruses BK virus (BKV) and JC virus (JCV) during childhood. After initial infection both viruses persist within renal system. Reactivation of both viruses may be linked with immunodeficiency or immunosuppressive therapy.


OBJECTIVE
To evaluate the relationship between immunodeficiency and viruria, prevalence of BK and JC viruria over time was investigated in a cohort of HIV seropositive individuals at different stages of disease. The excretion in this group was compared with virus excretion in their HIV seronegative partners and in an unselected cohort of patients attending a Genito-Urinary Medicine (GUM) clinic.


METHODS
The excretion of BKV and JCV DNA in multiple urine samples from HIV-infected patients at different stages of disease and their HIV-negative partners, and in single samples from a cohort of patients at a GUM clinic was investigated. A microplate hybridisation method was developed to increase both the sensitivity and specificity of detection of the PCR product. The method was also applied to estimate the DNA copy numbers of BKV and JCV in urine samples.


RESULTS
Within the HIV group, the level of immunosuppression (CD4+ category) was not associated with JCV viruria. By contrast, there was a modest correlation between immunodeficiency as indicated by a decline in CD4+ count and BKV viruria. Shedding of both BKV and JCV DNA together in urine samples of HIV-infected patients was much higher than in control groups (P = 0.02), indicating that HIV infection may associate with polyomavirus reactivation. The incidence of flu-like syndrome was much higher in HIV-infected asymptomatic individuals than acquired immunodeficiency syndrome (AIDS)-related complex (ARC)/AIDS patients. In general, the concentration of BKV DNA viruria (DNA copy number) was dependent to CD4+ counts (P = 0.008) while concentration of JCV DNA was independent to CD4+ cell count (P = 0.54). The prevalence of BKV and JCV DNA in patients who were infected with C. trachomatis was 9/50 (18%) and 11/50 (22%), respectively. BKV and JCV DNA was detected in 3/19 (15%) and 2/19 (10%) of patients who were infected with N. gonorrhoea. Results suggested that persons infected with C. trachomatis were more likely to show BKV and JCV viruria.


CONCLUSIONS
These results confirm that shedding of BK and JC viruses in urine is not exclusively found in immunosupression, it may also occur in healthy individuals. The frequency of virus excretion is however, apparently increased in HIV-infected patients, although no firm statistical difference could be established. One of the interesting aspects of these findings was the relatively high incidence of BKV and JCV viruria in both control groups, i.e. HIV-negative partners of HIV-infected patients and patients attending a GUM clinic.
Behzad-Behbahani
A
A
Clinical Virology Section, Organ Transplant Research Center (OTRC), Shiraz University of Medical Sciences, Shiraz, Iran. behbahani_2000@yahoo.com
Klapper
P E
PE
Vallely
P J
PJ
Cleator
G M
GM
Khoo
S H
SH
D000293:Adolescent; D000328:Adult; D000368:Aged; D001739:BK Virus / Q000235:genetics / Q000302:isolation & purification*; D018791:CD4 Lymphocyte Count; D004279:DNA, Viral / Q000652:urine; D005260:Female; D015658:HIV Infections / Q000821:virology*; D006801:Humans; D007121:Immunocompetence; D016867:Immunocompromised Host; D007577:JC Virus / Q000235:genetics / Q000302:isolation & purification*; D008297:Male; D008875:Middle Aged; D027601:Polyomavirus Infections / Q000821:virology*; D014556:Urine / Q000821:virology*; D019562:Viral Load; D014775:Virus Activation; D017201:Virus Shedding
D004279:DNA, Viral