Akademik Veri Yönetim Sistemi
Turk Hijyen ve Deneysel Biyoloji Dergisi, cilt.72, sa.4, ss.297-302, 2015 (Scopus)
Objective: BKV is a virus usually undergone asymptomatically in the childhood and can remain latent in the peripheral blood, brain and especially in kidneys. Reactivation of BKV under immunosuppression can cause diseases like interstitial nephritis, haemorrhagic or non- haemorrhagic cystitis, ureterostenosis and nephropathy. Especially in transplant recipients nephropathy frequency can reach 5% and can be the cause of premature loss 30-60% of transplanted organs and poor outcome. Quantification of BKV viral load in urine and serum with real time polymerase chain reaction (PCR) plays important role for early diagnosis and management of the therapy. Since the Real time PCR assay is more sensitive than classical PCR, can do quantification and have a less risk of contamination and short turn-around time. The aim of our study was to evaluate the analytical performance of a real time quantitative BKV PCR assay which was developed in our laboratory. Method: Standards were prepared from BKV plasmid (ATCC 45025). BKV plasmid that contained 15 x 107 copies/ ml to 3 x 101 copies/ml serial dilutions was measured by spectrofotometery. Primers for BKV VP1 gene and dual labelled probe at the 5' end with 6-carboxyfluoresceine (FAM) and the 3' end with 6-carboxytetramethylrhodamine (TAMRA) as described previously were used for the amplification reactions. Results: To evaluate the analytical performance of the assay; analytical sensitivity, specificity, linearity, accuracy and precision was determined. The analytical sensitivity and the limit of detection of the assay were found 15 x 102 copies/ml and 5 x 102 copies/ ml, respectively. Standard deviation (SD) of dilutions varied from 0.02 to 0.644 and CV varied from 0.79% to 11.47% between 15 x 107 to 15 x 101 copies/ml concentrations. Nineteen proficiency samples results of quality control program were in close agreement (100%). The assay demonstrated a linear range from 15 x 102 to 15 x 108 copies/ml. Specificity of the assay was found 100%. In addition proficiency samples results of the external quality control program were in close agreement. Conclusion: According to our results the real time PCR protocol of BKV developed in our laboratory was found sensitive, specific, precise and reproducible with a broad dynamic range.