Abstract
The quality of fecal specimens is one of the factors responsible for successful Clostridioides difficile infection (CDI) diagnosis. The quality depends largely on the storage conditions, including the temperature and time period. In this study, we organized the outputs of previous studies, filled experimental gaps in the knowledge of storage conditions, and introduced a pragmatic strategy for fecal storage for CDI diagnosis. A 5-step pathway was adopted to develop the fecal specimen storage strategy as follows: step 1, bibliomic analysis; step 2, experimental gap-filling; step 3, comparative evaluation; step 4, strategy development; step 5, internal review. Step 1 identified eight articles providing experimental information on the effects of fecal specimen storage conditions on the effectiveness of C. difficile detection methods. Step 2 provided additional quantitative data on C. difficile vegetative and spore cell viability and DNA stability. All previous and current results were compared (step 3). In step 4, fir general and nine special strategies were developed, followed by an internal review of the overall approaches (step 5). It is recommended to separate fecal samples into aliquots before testing and storing them. It is particularly recommended that fecal specimen samples be stored for CDI diagnosis at 4 °C for up to 60 days for all test methods.
Highlights
A Gram-positive, spore-forming, and obligate anaerobic bacterium, Clostridioides difficile, is responsible for the majority of recently increasing cases of infectious antibiotic-associated diarrhea and pseudomembranous colitis [1,2,3]
Several studies have investigated the impact of different storage conditions on the quality of stool samples [16,17,18,19,20,21,22,23]; these previous studies have some limitations that minimize their utility and there were experimental gaps in the storage conditions tested in the studies, preventing pragmatic storage strategies from being generated for stand-alone tests or currently accepted C. difficile infection (CDI)
We examined the differences in the numbers of C. difficile vegetative and spore cells during storage at −70 ◦ C, −20 ◦ C, 4 ◦ C, and RT over 28 days, which are typical storage temperature conditions
Summary
A Gram-positive, spore-forming, and obligate anaerobic bacterium, Clostridioides difficile, is responsible for the majority of recently increasing cases of infectious antibiotic-associated diarrhea and pseudomembranous colitis [1,2,3]. A cell culture cytotoxicity assay has been widely used for CDI diagnosis due to its high sensitivity, together with toxigenic culture [9,10]. Many laboratories use stand-alone tests or diagnostic algorithms to aid in the diagnosis of CDI [15]. While each of these test methods or diagnostic algorithms have their own benefits, the most critical factor to the accurate diagnosis of C. difficile is the quality of the fecal samples (and the corresponding targets of the test methods, e.g., cell viability, cytotoxicity, DNA stability, etc.). Several studies have investigated the impact of different storage conditions on the quality of stool samples [16,17,18,19,20,21,22,23]; these previous studies have some limitations that minimize their utility and there were experimental gaps in the storage conditions tested in the studies, preventing pragmatic storage strategies from being generated for stand-alone tests or currently accepted CDI diagnosis algorithms
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