%0 Journal Article
%T 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
%A Julu Bhatnagar
%A Marlene DeLeon-Carnes
%A Kathryn L. Kellar
%A Kakali Bandyopadhyay
%A Zoi-Anna Antoniadou
%A Wun-Ju Shieh
%A Christopher D. Paddock
%A Sherif R. Zaki
%J Infectious Diseases in Obstetrics and Gynecology
%D 2012
%I Hindawi Publishing Corporation
%R 10.1155/2012/972845
%X 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. 1. Introduction Clostridium species are ubiquitous Gram-positive, anaerobic, spore-forming bacteria that are generally found in soil and in the intestinal tract of humans and other animals. Clostridium sordellii has been reported to cause a variety of diseases including peritonitis, endocarditis, pneumonia, arthritis, cellulitis, and myonecrosis [1¨C4]. Fulminant toxic shock syndrome and sepsis among previously healthy persons have been described most often in cases associated with gynecologic infections and neonatal omphalitis [3, 5]. Clostridium perfringens is also responsible for a number of clinical conditions in humans ranging from acute food poisoning and enteritis to gas gangrene, enterotoxemia, and endometritis [6¨C8]. In the last couple of years, several studies reported pregnancy-associated fatal toxic shock syndrome cases due to C. sordellii and C. perfringens infections [8¨C10]. Among all C. sordellii infections reported in the literature, the overall case fatality ratio is 70%, while, for obstetric infections, it is more than 90% [8]. Pregnancy-associated C. perfringens fulminant septicemia also carries a very high mortality rate [8]. Deaths from C. sordellii and C. perfringens toxic shock
%U http://www.hindawi.com/journals/idog/2012/972845/