Evaluation of three enzyme immunoassays and a loop-mediated isothermal amplification test for the laboratory diagnosis of Clostridium difficile infection

Bruins, Marjan J.; Verbeek, E.; Wallinga, J.A.; Coppenraet, L.E.S. Bruijnesteijn van; Kuijper, Ed J.; Bloembergen, Peter

doi:10.1007/s10096-012-1658-y

Cited by 29 publications

(26 citation statements)

References 31 publications

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“…The early accurate identification of toxigenic C. difficile helps to direct resources and measures to appropriate targets. The development of nucleic acid amplification assays has recently enabled significant improvements in the detection of toxigenic C. difficile isolates in fecal specimens (14,19,23,24,27,36). Some of these methods, however, involve multiple manual or automated sample preparation steps, rendering the tests laborious and/or time-consuming.…”

Section: Discussionmentioning

confidence: 99%

“…Molecular assays are more sensitive than IA tests or EIAs and in some studies are even more sensitive than cytotoxigenic cultures or cytotoxin assays (21,22). Some of these methods have been commercialized and made suitable for use in clinical microbiological laboratories (14,(23)(24)(25)(26)(27). The main advantage of molecular assays, in addition to their high sensitivity and specificity, is the short turnaround time, compared with cytotoxigenic cultures or cytotoxin assays.…”

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confidence: 99%

“…These are sensitive but have several drawbacks, including long turnaround times and labor-intensive sample preparation and analysis, which limit their clinical utility (9)(10)(11)(12)(13). An alternative approach is the detection of C. difficile toxins or glutamate dehydrogenase (GDH) in stool samples by immunochromatographic antigen (IA) tests or enzyme immunoassays (EIAs) (14)(15)(16). These assays are rapid but are less sensitive and less specific than toxigenic cultures.…”

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confidence: 99%

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Evaluation of a New Automated Homogeneous PCR Assay, GenomEra C. difficile, for Rapid Detection of Toxigenic Clostridium difficile in Fecal Specimens

2013

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bWe evaluated a new automated homogeneous PCR assay to detect toxigenic Clostridium difficile, the GenomEra C. difficile assay (Abacus Diagnostica, Finland), with 310 diarrheal stool specimens and with a collection of 33 known clostridial and nonclostridial isolates. Results were compared with toxigenic culture results, with discrepancies being resolved by the GeneXpert C. difficile PCR assay (Cepheid). Among the 80 toxigenic culture-positive or GeneXpert C. difficile assay-positive fecal specimens, 79 were also positive with the GenomEra C. difficile assay. Additionally, one specimen was positive with the GenomEra assay but negative with the confirmatory methods. Thus, the sensitivity and specificity were 98.8% and 99.6%, respectively. With the culture collection, no false-positive or -negative results were observed. The analytical sensitivity of the GenomEra C. difficile assay was approximately 5 CFU per PCR test. The short hands-on (<5 min for 1 to 4 samples) and total turnaround (<1 h) times, together with the high positive and negative predictive values (98.8% and 99.6%, respectively), make the GenomEra C. difficile assay an excellent option for toxigenic C. difficile detection in fecal specimens.T oxin-producing Clostridium difficile is the most common cause of hospital-acquired and antibiotic-associated diarrhea (1, 2). The main virulence factors of C. difficile are C. difficile toxin A (TcdA) and C. difficile toxin B (TcdB), which are encoded by the genes tcdA and tcdB, respectively (3, 4). In addition, a separate binary toxin is produced by a small group of isolates with or without TcdA and/or TcdB (5, 6), and it has been suggested that the binary toxin may play a part in the recurrence of C. difficile infection (CDI) (7). As the role of the binary toxin in CDI currently is not well understood, it has not become a common target for diagnostic assays.CDI is prevalent in many hospitals and health care facilities throughout the world. It is associated with increases in hospitalization times and mortality rates, leading to augmented health care costs (2,6,8). In order to limit the transmission of C. difficile and to decrease the burden of CDI, rapid reliable detection of toxigenic C. difficile isolates and good infection prevention practices are required.Cytotoxigenic cultures and cytotoxin assays are considered standard diagnostic methods. These are sensitive but have several drawbacks, including long turnaround times and labor-intensive sample preparation and analysis, which limit their clinical utility (9-13). An alternative approach is the detection of C. difficile toxins or glutamate dehydrogenase (GDH) in stool samples by immunochromatographic antigen (IA) tests or enzyme immunoassays (EIAs) (14-16). These assays are rapid but are less sensitive and less specific than toxigenic cultures. Moreover, relying only on GDH detection reveals nothing about the toxigenic nature of the possible C. difficile isolates. To improve the detection of CDI using IA tests or EIAs, a two-step diagnostic algorithm has bee...

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

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Evaluation of a New Automated Homogeneous PCR Assay, GenomEra C. difficile, for Rapid Detection of Toxigenic Clostridium difficile in Fecal Specimens

2013

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“…After reviewing the titles and abstracts, 21 articles were selected for full-text evaluation (Figure 1). Sixteen articles with 18 studies published between 2005 and 2014 reported the sensitivity and specificity of LAMP on stool samples for the diagnosis of C. difficile infection, and were included in our meta-analysis [22][23][24][25][26][27][28][29][30][31][32][33][34][35][36][37]. Five articles were excluded (reasons for exclusion in Figure 1).…”

Section: Study Characteristicsmentioning

confidence: 99%

Accuracy of loop-mediated isothermal amplification for the diagnosis of Clostridium difficile infection: a systematic review

Lloyd

Pasupuleti

Thota

et al. 2015

Diagnostic Microbiology and Infectious Disease

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CitationLloyd A, Pasupuleti V, Thota P, Pant C, Rolston DDK, Hernandez A V., et al. This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. A C C E P T E D M A N U S C R I P T ACCEPTED MANUSCRIPT

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“…30-60 dakikada oluşan renk değişikliği basit bir türbidimetre ile veya çıplak gözle tespit edilebilmektedir. Illumigene yönteminin duyarlılığı %93.3-95.2, özgüllüğü ise %96.6-99.7 arasında değişmektedir 33,34 .…”

Section: Nükleik Asit Amplifikasyon Yöntemleriunclassified

Clostridium difficile Infection: Epidemiology, Risk Factors, Pathogenesis, Clinical Features, Diagnosis and Therapy

Kılıç

2013

Mikrobiyol Bul

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ÖZETClostridium difficile gram-pozitif, spor oluşturan anaerobik bir bakteridir. C.difficile antibiyotik ile ilişki-li ishal ve kolitlerin en önemli nedenidir. C.difficile enfeksiyonu (CDE) klinik spektrumu, hafif ishalden toksik megakolon, ileus, bağırsak perforasyonu ve psödomembranöz koliti içeren şiddetli intestinal hastalık tablosuna kadar oldukça değişken seyretmektedir. İleri yaş, hastanede kalış süresi ve özgün antibiyotiklere maruz kalma CDE için en yaygın risk faktörleridir. C.difficile'nin temel virülans determinantları toksin A (enterotoksin) ve toksin B (sitotoksin) olup, CDE tanısı dışkı örneklerinde C.difficile toksin A ve/veya toksin B'nin tespit edilmesine dayanmaktadır. C.difficile ve toksinleri tespit etmek için günümüzde çeşitli laboratuvar testleri geliştirilmiştir. Hücre kültürü sitotoksisite ve toksijenik kültür yöntemleri C.difficile toksinlerinin tespitinde referans yöntem olarak kabul edilmektedir. Ancak her iki yöntem de uygulanması güç, zaman alıcı ve uzman personel gerektiren yöntemlerdir. Bu nedenle çoğu mikrobiyoloji laboratuvarında daha pratik ve hızlı olan enzim temelli immünolojik yöntemler, glutamat dehidrojenaz antijen testleri ve gerçek zamanlı polimeraz zincir reaksiyonu (PCR) kullanılmaktadır. CDE tedavisinde ilk seçilecek antibiyotikler metronidazol ve vankomisindir. Bu derleme yazıda, CDE epidemiyolojisi, klinik özellikleri, risk faktörleri, patojenitesi, tanı testleri ve tedavisi özetlenmiştir. ABSTRACTClostridium difficile is a gram-positive, spore-forming anaerobic bacterium. C.difficile is the leading cause of antibiotic associated diarrhea and colitis. The clinical spectrum of C.difficile infection (CDI) is highly variable, ranging from mild diarrhea to severe forms of intestinal illness including toxic megacolon, ileus, bowel perforation, and pseudomembranous colitis. Advanced age, long duration of hospitalization, and exposure to certain antimicrobial agents are the most common risk factors for CDI. The main virulence determinants of C.difficile are toxin A (enterotoxin) and toxin B (cytotoxin). Diagnosis of CDI is based on the identification of C.difficile toxin A or B in diarrheal stool. Various laboratory tests have been currently developed for the detection of C.difficile or its toxins in stool samples. The cell culture cytotoxicity assay and toxigenic culture have been regarded as the reference standard methods for the detection of C.difficile toxins. However, both of the reference methods are laborious, time consuming, and need expert personnel. Therefore, many microbiology laboratories use enzyme immunoassays, glutamate dehydrogenase antigen tests and real-time polymerase chain reaction (PCR) which are more rapid and practical. First-line antibiotics for CDI treatment are metronidazole and vancomycin. In this review, epidemiology, clinical spectrum, risk factors, pathogenesis, diagnostic methods and treatment of CDI have been summarized.

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Evaluation of three enzyme immunoassays and a loop-mediated isothermal amplification test for the laboratory diagnosis of Clostridium difficile infection

Cited by 29 publications

References 31 publications

Evaluation of a New Automated Homogeneous PCR Assay, GenomEra C. difficile, for Rapid Detection of Toxigenic Clostridium difficile in Fecal Specimens

Evaluation of a New Automated Homogeneous PCR Assay, GenomEra C. difficile, for Rapid Detection of Toxigenic Clostridium difficile in Fecal Specimens

Accuracy of loop-mediated isothermal amplification for the diagnosis of Clostridium difficile infection: a systematic review

Clostridium difficile Infection: Epidemiology, Risk Factors, Pathogenesis, Clinical Features, Diagnosis and Therapy

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