What Pre-FecalSwab or Molecular Methods for Detecting Enteric Pathogens Look Like™
Traditionally, detecting pathogens in stool happens in one of two ways. Microscopy is an option, which can be troublesome as that it is both subjective and requires a high amount of expertise from the technologist. The other method is culture based identification of bacterial pathogens, which involves plating the specimen for growth on various selective media, followed by additional biochemical/phenotypic testing for confirmation. Therefore, in a situation where one needs a quick and reliable identification, resources are either limited or confirmation requires extra time. Dynacare Microbiology Laboratories are a high-volume laboratory located in Ontario, Canada that receives over 200 specimens a day that call for enteric diagnosis. As is the case with most laboratories with high volumes, finding an “accurate diagnostic method with efficient workflow” becomes a high priority. Thus, Seegene’s Allplex Gastrointestinal Multiplex PCR and FecalSwab™ were investigated as an option for fast results to enteric investigations and efficient workflow. FecalSwab™ Versus Traditional Enteric Cary-Blair Transport Systems A unique FLOQSwab™, paired with 2 mL of Cary-Blair in compact plastic tubes in COPAN’s fecal collection and transport kit provide the ideal ratio of fecal matter to
transport media for easier use with the molecular assays. The evaluation of FecalSwab™ compared to traditional Cary-Blair transport systems involved: Bacterial Enteric Pathogens via culture, Bacterial Enteric Pathogens via molecular testing and Protozoan Enteric Pathogens via molecular testing. Demonstrated in the study, the complete value chain of activities was assessed. First the investigators looked at the preanalytical phase, which includes the specimen collection, transportation, and set up for DNA extraction and PCR plates. Next, they examined the analytical phase, which includes the actual PCR, reflex test setup for culture and antibiotic susceptibility testing. Finally, the postanalytical phase consisting of the reporting and referral for confirmation testing was scrutinized.
FecalSwab™ for Use with Seegene Allplex Gastrointestinal Multiplex PCR
The overall process for using FecalSwab™ with the Seegene Allplex gastrointestinal multiplex PCR is simple. Collect using the FLOQSwab™ included in the FecalSwab™ kit by slightly dipping it into the stool sample, like a probe and loading it into the tube,
careful not to overfill past the marked “max fill” line indicated on the tube. Mash and mix the stool until the sample appears homogenous and recap the tube. Once ready, uncap the fecal transport kit tubes and place directly onto the liquid handling platforms in the lab, from there the system does DNA extraction and handles the Master mix preparation. The fully prepared Seegene assay plate is then transferred to real-time thermocycler for analysis. It is important to note that in this study FecalSwab was used in conjunction with two different liquid handling platforms, due to their use in the lab at Dynacare, and include the Hamilton STARLet and the Hamilton Nimbus. Among some of the key benefits they observed by using COPAN’s FecalSwab is that it demonstrated an improved limit of detection for both culture and molecular testing! To begin, FecalSwab™ specimens were simply de-capped and loaded directly onto the Hamilton Nimbus or Starlet platforms resulting in higher productivity from an operational standpoint and improved the audit trail during automated DNA extraction and PCR plate setup. The sensitivity to detecting bacterial pathogens including Salmonella sp, Shigella sp, Campylobacter jejuni/coli, Yersinia enterocolitica and E. coli O157 was also improved dramatically, between 10 – 100-fold, compared to traditional Cary-Blair Enteric transport media. In addition, with the ability to test for both parasites and bacteria from a single sample, not only were the transportation and pre-analytical processes streamlined, but the analytical sensitivity was also enhanced. The optimized design and Cary-Blair formulation of FecalSwab™ allows for traditional reflex bacteriological culture, antibiotic susceptibility and epidemiological typing. Whether its automated liquid handling platforms or molecular assays for enteric pathogens for bacteria, viruses or parasites, FecalSwab™ outperforms traditional enteric transport media. FecalSwab™ Improves Detection with Less Sample The results of the study are promising and show that using FecalSwab™ significantly improves quality when it comes to detecting enteric pathogens, even if a molecular test is not administered. The numbers showed viability of enteric bacterial pathogens are improved in the COPAN FecalSwab™, permitting growth at lower dilutions. In addition, the stability of enteric bacterial pathogens was increased as PCR detection was occurring at lower dilution. PCR detection was possible at lower dilutions with the FecalSwab™ than with the traditional Cary-Blair sample transports for Protozoan Parasitic pathogens, as well! When samples were placed into FecalSwab™, lower Ct values for parasite detection was observed, enhancing PCR detection, as well asdetection of Parasites at lower dilutions!!
The successful examination of COPAN’s FecalSwab™ and its compatible use with yet another gastrointestinal multiplex PCR is testimonial not only to the integrity and versatility of the product, but also the widespread adoption of its use among clinicians, microbiologists and assay manufacturers alike. The reason for its success? FecalSwab™ works. When there’s an opportunity for improvement, COPAN seizes it. Rather than waiting for someone else to create the solution, products are developed with excellence in mind, because at the end of every product there is a patient who not only deserves quality, but depends on it.
With a reputation for innovation, COPAN is a worldwide leader in the manufacturing of collection and transport systems. Copan’s collaborative approach to preanalytics has resulted in Flocked Swabs, ESwab, Universal Transport Medium and laboratory automation, WASP® and WASPLab™. COPAN’s collection and systems have been proven to advance the quality of traditional and contemporary microbiology assays. COPAN's automation includes specimen processing, smart incubation, digital imaging, and strong algorithms for automatic segregation of bacterial cultures, followed by automated colony picking. COPAN carries a wide range of environmental sampling products for the food, beverage, hospital and pharmaceutical industries. For more information, visit http://www.copanusa.com/