Test: Babesia DNA qPCR on Whole Blood Samples
Real time polymerase chain reaction (PCR) is one of the many variants of PCR used in molecular diagnostics. Real time PCR represents a sensitive and powerful tool for diagnosis and monitoring of acute and chronic infectious agents in patient samples.
This procedure detects the presence of Babesia 18S rRNA genes in human whole blood. Total nucleic acids are purified from the sample. Conserved sequences of Babesia genes of interest are amplified using genus-specific primers. The presence of amplicon allows a TaqMan probe to bind and be cleaved during replication. As amplicon quantity increases, the fluorescent signal increases. If the sample is positive the fluorescent signal will pass a pre-determined threshold. The cycle at which the fluorescence passes the threshold is the cycle threshold (Ct). The Ct value is compared to a standard curve of synthetic double-stranded DNA (DSD) Ct values to determine a relative quantity of Babesia DNA in the sample. If no amplicon is created, then no probe will be cleaved providing no fluorescent signal. Calibrator, medium positive, low positive and negative control reactions are performed with each test.
Volume / Collection Type
Whole blood6 – 10 mL collected in ACD tube. EDTA also acceptable3 mLRoom temperature for < 48 hoursRoom temperature
Specimen:Whole bloodVolume / Collection Type:6 – 10 mL collected in ACD tube. EDTA also acceptableMinimum:3 mLStorage:Room temperature for < 48 hoursShipping:Room temperatureNote: Specimens must be received by lab before expiration of storage times.
Turn Around Time
Results available within four business days of specimen receipt in the lab.
Reported as copies/mL and log copies/mL
All specimens must be labeled with the patient’s name and collection date. Specimens are accepted Monday through Friday NOON (Central). Call for prior arrangements beyond these accepted times.