A Novel Subtyping Assay for Detection of Clostridium difficile Virulence Genes

• Published in: The Journal of molecular diagnostics : JMD Vol. 16; no. 2; pp. 244 - 252
• Main Authors: Angione, Stephanie L., Sarma, Aartik A., Novikov, Aleksey, Seward, Leah, Fieber, Jennifer H., Mermel, Leonard A., Tripathi, Anubhav
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.03.2014; American Society for Investigative Pathology
• Subjects: ADP Ribose Transferases - genetics; Bacterial Proteins - genetics; Bacterial Toxins - genetics; Clostridium difficile - genetics; Enterocolitis, Pseudomembranous - diagnosis; Humans; Multiplex Polymerase Chain Reaction - methods; Pathology; Point-of-Care Systems; Real-Time Polymerase Chain Reaction - methods; Regular; Reproducibility of Results; Sensitivity and Specificity; Virulence - genetics; Virulence Factors - genetics
• ISSN: 1525-1578; 1943-7811; 1943-7811
• DOI: 10.1016/j.jmoldx.2013.11.006

This proof-of-concept study demonstrates the application of a novel nucleic acid detection platform to detect Clostridium difficile in subjects presenting with acute diarrheal symptoms. This method amplifies three genes associated with C. difficile infection, including genes and deletions (cdtB and tcdC) associated with hypervirulence attributed to the NAP1/027/BI strain. Amplification of DNA from the tcdB, tcdC, and cdtB genes was performed using a droplet-based sandwich platform with quantitative real-time PCR in microliter droplets to detect and identify the amplified fragments of DNA. The device and identification system are simple in design and can be integrated as a point-of-care test to help rapidly detect and identify C. difficile strains that pose significant health threats in hospitals and other health-care communities.