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Detection of Clostridium difficile toxin in various tissue culture monolayers
A C Maniar1, T W Williams, G W Hammond
1Cadham Provincial Laboratory, Winnipeg, Manitoba, Canada.
Abstract:
Thirty stool filtrates known to contain Clostridium difficile toxin based on previous testing on McCoy cells were tested for toxicity on primary African green monkey kidney (AGMK), McCoy, MRC-5, primary rhesus monkey kidney (RMK), and Vero cells. All 30 filtrates showed cytotoxic effect at greater than or equal to 1:100 dilution on McCoy and Vero cells. A total of 22 filtrates were positive on MRC-5 monolayers, while only 16 and 10 filtrates showed positive cytotoxic effect on AGMK and RMK cells, respectively. Another 630 stool specimens were tested on McCoy and Vero cells only. Of these stool filtrates, 70 were positive and 560 were negative with both cell lines, which thus gave 100% agreement. Vero cells can be used interchangeably with McCoy cells for the detection of C. difficile toxin in stool filtrates.
Insights
Vero cells and McCoy cells are equally effective for detecting Clostridium difficile toxin in stool filtrates. This finding supports using Vero cells as a reliable alternative for C. difficile toxin testing.
Area of Science:
- Microbiology
- Cell Biology
- Toxicology
Background:
- Clostridium difficile infection (CDI) is a significant cause of antibiotic-associated diarrhea.
- Accurate and efficient detection of C. difficile toxin is crucial for timely diagnosis and treatment.
- Traditional cell-based assays for toxin detection have been used, but alternative cell lines are continually evaluated.
Purpose of the Study:
- To evaluate the efficacy of Vero cells compared to McCoy cells and other cell lines for detecting Clostridium difficile toxin in stool filtrates.
- To determine if Vero cells can serve as a reliable substitute for McCoy cells in routine diagnostic testing.
Main Methods:
- Stool filtrates previously identified as containing C. difficile toxin were tested for cytotoxicity on multiple cell lines: African green monkey kidney (AGMK), McCoy, MRC-5, rhesus monkey kidney (RMK), and Vero cells.
- Cytotoxicity was assessed based on the observed effects at specific dilutions.
- A larger set of stool specimens (630) was subsequently tested on McCoy and Vero cells to confirm agreement.
Main Results:
- All 30 tested stool filtrates demonstrated cytotoxic effects at a dilution of 1:100 or greater on both McCoy and Vero cells.
- MRC-5 cells detected toxin in 22 filtrates, while AGMK and RMK cells were positive for 16 and 10 filtrates, respectively.
- Testing of 630 additional stool specimens showed 100% agreement between McCoy and Vero cells, with 70 positive and 560 negative results.
Conclusions:
- Vero cells exhibit comparable sensitivity and specificity to McCoy cells for the detection of C. difficile toxin.
- Vero cells are a suitable and interchangeable alternative to McCoy cells for C. difficile toxin testing in stool specimens.
- The use of Vero cells can streamline diagnostic workflows for C. difficile infections.