Mycoplasma DNA Detection Kit (qPCR), BP-QN22

qPCR Sensitivity Verification

Table 1. Summary of Limit-of-Detection Data for 10 Common Standard Mycoplasma Strains

At 5 CFU/mL, the kit detected every standard Mycoplasma strain in 24 of 24 tests.

Cross-Reactivity Results

Cross-reactivity was evaluated using structurally related microorganisms and commonly used cell lines. DNA from 27 panel-listed items (CHO, Vero, E. coli, Sf9, MDCK, HEK293T, SP2/0, MDBK, PK15, CRFK, LLC-MK2, C6, MRC-5, Pichia pastoris, Lactobacillus acidophilus, Candida albicans, Acinetobacter baumannii, Pseudomonas aeruginosa, Staphylococcus epidermidis, Streptococcus pneumoniae, Staphylococcus aureus, Streptococcus mutans, Monascus purpureus, Salmonella enterica, Staphylococcus intermedius, Enterobacter aerogenes, and Streptococcus agalactiae) showed no cross-reactivity with the Mycoplasma detection reagent. The results are shown in Table 2.

Table 2. Cross-Reactivity Results

Conclusion:

DNA from the 27 panel-listed items (CHO, Vero, E. coli, Sf9, MDCK, HEK293T, SP2/0, MDBK, PK15, CRFK, LLC-MK2, C6, MRC-5, P. pastoris, L. acidophilus, C. albicans, A. baumannii, P. aeruginosa, S. epidermidis, S. pneumoniae, S. aureus, S. mutans, M. purpureus, S. enterica, S. intermedius, E. aerogenes, and S. agalactiae) showed no cross-reactivity with the Mycoplasma detection reagent.

Bioinformatics Analysis

Sequence alignment of the amplification target against the NCBI database indicated coverage of more than 200 Mycoplasma species.

Precision

Within-Run Precision

In-house reference materials Z1, Z2, and Z3 were tested in 10 replicates each. Precision CV was within 5% at every concentration.

Intermediate Precision

Intermediate precision was evaluated across operators, days, instruments, and reagent lots using in-house reference materials Z1, Z2, and Z3, with five replicates per material. Precision CV was within 5% at every concentration.

Total Imprecision

Data across operators, days, instruments, and reagent lots were pooled. Precision CV was within 5% at every concentration.

Performance data shown here were obtained with the reference materials, extraction workflows, instruments and acceptance criteria used in BIORI’s product-specific validation study. Users should verify performance in their own sample matrix and workflow.