Comparison of Microdroplet Digital PCR Assays with Real-time Fluorescence Quantitative PCR for Clostridioides difficile Detection

Clostridioides difficile(C.difficile),is an anaerobic Gram-positive bacterium found in the human intestine.Prolonged use of antibacterial drugs can result in C.difficile infection(CDI)[1],which is one of the most important hospital-acquired diseases affecting all wards.Isolation-and-culture-based identification and the cell cytotoxicity neutralization assay(CCNA)are considered the“gold standard”for detecting toxins or pathogenic strains[2],with long and time-consuming incubation times.Fluorescent quantitative polymerase chain reaction(PCR)is commonly used in clinical laboratory tests[3].However,the sensitivity,accuracy,and resolution of detection are limited by low-copy target gene molecules and subtle differences in template concentration.As C.difficile is distributed in feces in the natural environment and the sample matrix is complex,it is difficult to detect pathogens at low levels.Accurately quantifying C.difficile and implementing effective measures to combat C.difficile infections are of utmost importance.