Unknown

Dataset Information

0

Propidium Monoazide (PMAxx)-Recombinase Polymerase Amplification Exo (RPA Exo) Assay for Rapid Detection of Burkholderia cepacia Complex in Chlorhexidine Gluconate (CHX) and Benzalkonium Chloride (BZK) Solutions.


ABSTRACT: Both sterile and non-sterile pharmaceutical products, which include antiseptics, have been recalled due to Burkholderia cepacia complex (BCC) contamination. Therefore, minimizing the frequency of outbreaks may be conducive to the development of a quick and sensitive approach that can distinguish between live and dead loads of BCC. We have assessed an exo probe-based recombinase polymerase amplification (RPA) with 10 µM propidium monoazide (PMAxx) for selective detection of live/dead BCC cells in various concentrations of antiseptics (i.e., chlorhexidine gluconate (CHX) and benzalkonium chloride (BZK) solutions) after 24 h. The optimized assay conducted using a set of primer-probes targeting gbpT was performed at 40 °C for 20 min and shows a detection limit of 10 pg/µL of genomic DNA from B. cenocepacia J2315, equivalent to 104 colony-forming units (CFU/mL). The specificity of a newly designed primer and probe was 80% (20 negatives out of 25). The readings for total cells (i.e., without PMAxx) from 200 µg/mL CHX using PMAxx-RPA exo assay was 310 relative fluorescence units (RFU), compared to 129 RFU with PMAxx (i.e., live cells). Furthermore, in 50-500 µg/mL BZK-treated cells, a difference in the detection rate was observed between the PMAxx-RPA exo assay in live cells (130.4-459.3 RFU) and total cells (207.82-684.5 RFU). This study shows that the PMAxx-RPA exo assay appears to be a valid tool for the simple, rapid and presumptive detection of live BCC cells in antiseptics, thereby ensuring the quality and safety of pharmaceutical products.

SUBMITTER: Daddy Gaoh S 

PROVIDER: S-EPMC10302078 | biostudies-literature | 2023 May

REPOSITORIES: biostudies-literature

altmetric image

Publications

Propidium Monoazide (PMAxx)-Recombinase Polymerase Amplification Exo (RPA Exo) Assay for Rapid Detection of <i>Burkholderia cepacia</i> Complex in Chlorhexidine Gluconate (CHX) and Benzalkonium Chloride (BZK) Solutions.

Daddy Gaoh Soumana S   Kweon Ohgew O   Ahn Youngbeom Y  

Microorganisms 20230526 6


Both sterile and non-sterile pharmaceutical products, which include antiseptics, have been recalled due to <i>Burkholderia cepacia</i> complex (BCC) contamination. Therefore, minimizing the frequency of outbreaks may be conducive to the development of a quick and sensitive approach that can distinguish between live and dead loads of BCC. We have assessed an exo probe-based recombinase polymerase amplification (RPA) with 10 µM propidium monoazide (PMAxx) for selective detection of live/dead BCC c  ...[more]

Similar Datasets

| S-EPMC9147393 | biostudies-literature
| S-EPMC10882272 | biostudies-literature
| S-EPMC11822742 | biostudies-literature
| S-EPMC5120141 | biostudies-literature
| S-EPMC6680226 | biostudies-literature
| S-EPMC7819323 | biostudies-literature
2020-11-24 | GSE162029 | GEO
| S-EPMC11497490 | biostudies-literature
| S-EPMC7119533 | biostudies-literature
| S-EPMC8347691 | biostudies-literature