SO SÁNH HAI MASTER MIX PCR PROBE TRONG KỸ THUẬT REAL-TIME PCR XÁC ĐỊNH BK POLYOMAVIRUS

COMPARISON OF TWO MASTER MIXES REAL-TIME POLYMERASE CHAIN REACTION PROBE FOR BK POLYOMAVIRUS DETECTIONComparison of the efficiency of probe real-time PCR master mixes for BKV detection offers additional PCR reagent options for clinical laboratory tests. In the present study, AceQ qPCR Probe Master Mix was compared with QuantiTect Probe PCR Master Mix in the real-time PCR assay targeting the VP1-BKV gene. The Rotor-Gene Q platform (Qiagen, Germany) and a 10-fold serial dilution of recombinant BKV-VP1 plasmid ranging from 2.10-1 copies/µl to 2.105 copies/µl were used to evaluate. There were differences in Ct values (p<0.05) between the two master mixes but complete agreement in the performance of AceQ Probe MM compared with QuantiTect Probe MM, showing an excellent correlation with Pearson’s correlation coefficient (r) of 0.993 (p<0.0001). AceQ Probe MM as well as QuantiTect Probe MM showed good coefficient of determination (R2) values, high PCR efficiency (H%), and limit of detection at 95% confidence with R2 value of 0.994, 97.55%, LOD 95% of 0.87 (0.419-1.815) copies/µl, and R2 value of 0.991, 92.28%, LOD 1.17 (0.342-93.046) copies/µl, respectively. PCR assay running times of AceQ Probe MM and QuantiTect Probe MM were 1.17 and 1.65 hours, respectively. Furthermore, with clinical samples, Ct values comparison of all samples in which a positive result showed a high degree of linear regression analysis (r = 0.996) between the two master mixes when we evaluated eleven specimens, including plasma and urine samples from renal transplantation recipients. The Bland-Altman plot comparison showed that the average difference of -0.112 log10 BKV-DNA load, was concordance for BKV detection of two master mixes with all load differences in the range of the bias ± 1.96 SD (-0.299; 0.524). In conclusion, our study demonstrated that AceQ Probe MM and QuantiTect Probe MM were both suitable for real-time probe PCR BKV detection targeting the VP1 gene. The combination of the two probe PCR master mixes can be considered for rapid detection of BKV infection.