title: Rapid, simultaneous detection of Clostridium sordellii and Clostridium perfringens in archived tissues by a novel PCR-based microsphere assay: diagnostic implications for pregnancy-associated toxic shock syndrome cases.
abstract: Clostridium sordellii and Clostridium perfringens are infrequent human pathogens; however, the case-fatality rates for the infections are very high, particularly in obstetric C. sordellii infections (>90%). Deaths from Clostridium sordellii and Clostridium perfringens toxic shock (CTS) are sudden, and diagnosis is often challenging. Formalin-fixed, paraffin-embedded (FFPE) tissues usually are the only specimens available for sudden fatal cases, and immunohistochemistry (IHC) for Clostridia is generally performed but it cannot identify species. A clear need exists for a rapid, species-specific diagnostic assay for FFPE tissues. We developed a duplex PCR-based microsphere assay for simultaneous detection of C. sordellii and C. perfringens and evaluated DNA extracted from 42 Clostridium isolates and FFPE tissues of 28 patients with toxic shock/endometritis (20 CTS, 8 non-CTS, as confirmed by PCR and sequencing). The microsphere assay correctly identified C. sordellii and C. perfringens in all known isolates and in all CTS patients (10 C. sordellii, 8 C. perfringens, 2 both) and showed 100% concordance with PCR and sequencing results. The microsphere assay is a rapid, specific, and cost-effective method for the diagnosis of CTS and offers the advantage of simultaneous testing for C. sordellii and C. perfringens in FFPE tissues using a limited amount of DNA.
authors.affiliation: Infectious Diseases Pathology Branch, Division of High-Consequence Pathogens and Pathology, Centers for Disease Control and Prevention, Atlanta, GA 30333, USA. JBhatnagar@cdc.gov
authors.forename: Julu
authors.initials: J
authors.lastname: Bhatnagar
authors.forename: Marlene
authors.initials: M
authors.lastname: Deleon-Carnes
authors.forename: Kathryn L
authors.initials: KL
authors.lastname: Kellar
authors.forename: Kakali
authors.initials: K
authors.lastname: Bandyopadhyay
authors.forename: Zoi-Anna
authors.initials: ZA
authors.lastname: Antoniadou
authors.forename: Wun-Ju
authors.initials: WJ
authors.lastname: Shieh
authors.forename: Christopher D
authors.initials: CD
authors.lastname: Paddock
authors.forename: Sherif R
authors.initials: SR
authors.lastname: Zaki
mesh_terms: D000293:Adolescent; D000328:Adult; D003015:Clostridium Infections / Q000175:diagnosis*; D003016:Clostridium perfringens / Q000302:isolation & purification*; D047989:Paraclostridium sordellii / Q000302:isolation & purification*; D005260:Female; D006801:Humans; D008863:Microspheres; D016133:Polymerase Chain Reaction / Q000379:methods*; D011247:Pregnancy; D011251:Pregnancy Complications, Infectious / Q000175:diagnosis*; D012772:Shock, Septic / Q000175:diagnosis*; D055815:Young Adult