Supportive Data

The following validation data supports the use of this assay for clinical testing.

Accuracy/Diagnostic Sensitivity and Specificity:

Ninety-six whole blood specimens were tested by this real-time polymerase chain reaction (PCR) assay and another real-time PCR assay. Concordance was 99%.

Analytical Sensitivity/Limit of Detection:

The limit of detection established using whole organism spiked into specimen matrix (whole blood) is as follows:

-Babesia microti, ATCC PRA 99-2670 target copies/mL

-Babesia duncani ATCC PRA 302-1540 target copies/mL

-Babesia MO-1 positive patient DNA-10,700 target copies/mL

-Babesia divergens positive patient DNA-5270 target copies/mL

Serial 10-fold dilutions of microscopy-positive specimens were also tested in a blinded fashion using conventional thick and thin blood films and the Mayo Clinic Babesia species PCR test. The PCR test was able to consistently detect two 10-fold dilutions lower than using microscopy.

Analytical Specificity:

No cross-reactivity was noted using a panel of 34 bacteria, viruses, parasites, and fungi were detected by the Babesia species PCR.

Precision:

Interassay and intra-assay precision was 100% precision.

Reference Range:

The reference range is negative. This was confirmed by testing 93 blood specimens from asymptomatic individuals for the presence of Babesia species by the Babesia species PCR assay. All 93 specimens were negative.

Reportable Range:

This test is a qualitative assay, and results are reported as positive or negative for Babesia species (B microti, B duncani, B divergens, and Babesia MO-1).