Diagnosis of Clostridium difficile

real-time PCR detection of toxin genes in faecal samples is more sensitive compared to toxigenic culture

Mie Birgitte Frid Jensen, Katharina E. P. Olsen, Xiaohui Chen Nielsen, Anne Mette Hoeg, Ram Benny Dessau, Tove Atlung, Jørgen Engberg

    Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

    Resumé

    The diagnosis of Clostridium difficile infection (CDI) requires the detection of toxigenic C. difficile or its toxins and a clinical assessment. We evaluated the performance of four nucleic acid amplification tests (NAATs) detecting toxigenic C. difficile directly from faeces compared to routine toxigenic culture. In total, 300 faecal samples from Danish hospitalised patients with diarrhoea were included consecutively. Culture was performed in duplicate (routine and ‘expanded toxigenic culture’: prolonged and/or re-culture) and genotypic toxin profiling by polymerase chain reaction (PCR), PCR ribotyping and toxinotyping (TT) were performed on culture-positive samples. In parallel, the samples were analysed by four NAATs; two targeting tcdA or tcdB (illumigene® C. difficile and PCRFast® C. difficile A/B) and two multi-target real-time (RT) PCR assays also targeting cdt and tcdC alleles characteristic of epidemic and potentially more virulent PCR ribotypes 027, 066 and 078 (GeneXpert® C. difficile/Epi and an ‘in-house RT PCR’ two-step algorithm). The multi-target assays were significantly more sensitive compared to routine toxigenic culture (p < 0.05) and significantly more robust to inhibition compared to PCRFast (p < 0.001). Duplicate ‘expanded toxigenic culture’ increased the culture-positive rate by 29 % compared to routine culture. The ability of the GeneXpert and in-house assays to correctly classify PCR ribotype 027 was high (>95 %), and in-house PCR displayed 100 % correct identification of PCR ribotypes 066 and 078. Furthermore, the presence of the PCR enhancer bovine serum albumin (BSA) was found to be related to high sensitivity and low inhibition rate. Rapid laboratory diagnosis of toxigenic C. difficile by RT PCR was accurate
    OriginalsprogEngelsk
    TidsskriftEuropean Journal of Clinical Microbiology & Infectious Diseases
    Vol/bind34
    Udgave nummer4
    Sider (fra-til)727-736
    ISSN0934-9723
    DOI
    StatusUdgivet - apr. 2015

    Emneord

    • Clostridium difficile

    Citer dette

    Jensen, Mie Birgitte Frid ; Olsen, Katharina E. P. ; Nielsen, Xiaohui Chen ; Hoeg, Anne Mette ; Dessau, Ram Benny ; Atlung, Tove ; Engberg, Jørgen. / Diagnosis of Clostridium difficile : real-time PCR detection of toxin genes in faecal samples is more sensitive compared to toxigenic culture. I: European Journal of Clinical Microbiology & Infectious Diseases. 2015 ; Bind 34, Nr. 4. s. 727-736.
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    title = "Diagnosis of Clostridium difficile: real-time PCR detection of toxin genes in faecal samples is more sensitive compared to toxigenic culture",
    abstract = "The diagnosis of Clostridium difficile infection (CDI) requires the detection of toxigenic C. difficile or its toxins and a clinical assessment. We evaluated the performance of four nucleic acid amplification tests (NAATs) detecting toxigenic C. difficile directly from faeces compared to routine toxigenic culture. In total, 300 faecal samples from Danish hospitalised patients with diarrhoea were included consecutively. Culture was performed in duplicate (routine and ‘expanded toxigenic culture’: prolonged and/or re-culture) and genotypic toxin profiling by polymerase chain reaction (PCR), PCR ribotyping and toxinotyping (TT) were performed on culture-positive samples. In parallel, the samples were analysed by four NAATs; two targeting tcdA or tcdB (illumigene{\circledR} C. difficile and PCRFast{\circledR} C. difficile A/B) and two multi-target real-time (RT) PCR assays also targeting cdt and tcdC alleles characteristic of epidemic and potentially more virulent PCR ribotypes 027, 066 and 078 (GeneXpert{\circledR} C. difficile/Epi and an ‘in-house RT PCR’ two-step algorithm). The multi-target assays were significantly more sensitive compared to routine toxigenic culture (p < 0.05) and significantly more robust to inhibition compared to PCRFast (p < 0.001). Duplicate ‘expanded toxigenic culture’ increased the culture-positive rate by 29 {\%} compared to routine culture. The ability of the GeneXpert and in-house assays to correctly classify PCR ribotype 027 was high (>95 {\%}), and in-house PCR displayed 100 {\%} correct identification of PCR ribotypes 066 and 078. Furthermore, the presence of the PCR enhancer bovine serum albumin (BSA) was found to be related to high sensitivity and low inhibition rate. Rapid laboratory diagnosis of toxigenic C. difficile by RT PCR was accurate",
    keywords = "Clostridium difficile, Clostridium difficile",
    author = "Jensen, {Mie Birgitte Frid} and Olsen, {Katharina E. P.} and Nielsen, {Xiaohui Chen} and Hoeg, {Anne Mette} and Dessau, {Ram Benny} and Tove Atlung and J{\o}rgen Engberg",
    year = "2015",
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    doi = "10.1007/s10096-014-2284-7",
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    Diagnosis of Clostridium difficile : real-time PCR detection of toxin genes in faecal samples is more sensitive compared to toxigenic culture. / Jensen, Mie Birgitte Frid; Olsen, Katharina E. P.; Nielsen, Xiaohui Chen ; Hoeg, Anne Mette; Dessau, Ram Benny ; Atlung, Tove; Engberg, Jørgen.

    I: European Journal of Clinical Microbiology & Infectious Diseases, Bind 34, Nr. 4, 04.2015, s. 727-736.

    Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

    TY - JOUR

    T1 - Diagnosis of Clostridium difficile

    T2 - real-time PCR detection of toxin genes in faecal samples is more sensitive compared to toxigenic culture

    AU - Jensen, Mie Birgitte Frid

    AU - Olsen, Katharina E. P.

    AU - Nielsen, Xiaohui Chen

    AU - Hoeg, Anne Mette

    AU - Dessau, Ram Benny

    AU - Atlung, Tove

    AU - Engberg, Jørgen

    PY - 2015/4

    Y1 - 2015/4

    N2 - The diagnosis of Clostridium difficile infection (CDI) requires the detection of toxigenic C. difficile or its toxins and a clinical assessment. We evaluated the performance of four nucleic acid amplification tests (NAATs) detecting toxigenic C. difficile directly from faeces compared to routine toxigenic culture. In total, 300 faecal samples from Danish hospitalised patients with diarrhoea were included consecutively. Culture was performed in duplicate (routine and ‘expanded toxigenic culture’: prolonged and/or re-culture) and genotypic toxin profiling by polymerase chain reaction (PCR), PCR ribotyping and toxinotyping (TT) were performed on culture-positive samples. In parallel, the samples were analysed by four NAATs; two targeting tcdA or tcdB (illumigene® C. difficile and PCRFast® C. difficile A/B) and two multi-target real-time (RT) PCR assays also targeting cdt and tcdC alleles characteristic of epidemic and potentially more virulent PCR ribotypes 027, 066 and 078 (GeneXpert® C. difficile/Epi and an ‘in-house RT PCR’ two-step algorithm). The multi-target assays were significantly more sensitive compared to routine toxigenic culture (p < 0.05) and significantly more robust to inhibition compared to PCRFast (p < 0.001). Duplicate ‘expanded toxigenic culture’ increased the culture-positive rate by 29 % compared to routine culture. The ability of the GeneXpert and in-house assays to correctly classify PCR ribotype 027 was high (>95 %), and in-house PCR displayed 100 % correct identification of PCR ribotypes 066 and 078. Furthermore, the presence of the PCR enhancer bovine serum albumin (BSA) was found to be related to high sensitivity and low inhibition rate. Rapid laboratory diagnosis of toxigenic C. difficile by RT PCR was accurate

    AB - The diagnosis of Clostridium difficile infection (CDI) requires the detection of toxigenic C. difficile or its toxins and a clinical assessment. We evaluated the performance of four nucleic acid amplification tests (NAATs) detecting toxigenic C. difficile directly from faeces compared to routine toxigenic culture. In total, 300 faecal samples from Danish hospitalised patients with diarrhoea were included consecutively. Culture was performed in duplicate (routine and ‘expanded toxigenic culture’: prolonged and/or re-culture) and genotypic toxin profiling by polymerase chain reaction (PCR), PCR ribotyping and toxinotyping (TT) were performed on culture-positive samples. In parallel, the samples were analysed by four NAATs; two targeting tcdA or tcdB (illumigene® C. difficile and PCRFast® C. difficile A/B) and two multi-target real-time (RT) PCR assays also targeting cdt and tcdC alleles characteristic of epidemic and potentially more virulent PCR ribotypes 027, 066 and 078 (GeneXpert® C. difficile/Epi and an ‘in-house RT PCR’ two-step algorithm). The multi-target assays were significantly more sensitive compared to routine toxigenic culture (p < 0.05) and significantly more robust to inhibition compared to PCRFast (p < 0.001). Duplicate ‘expanded toxigenic culture’ increased the culture-positive rate by 29 % compared to routine culture. The ability of the GeneXpert and in-house assays to correctly classify PCR ribotype 027 was high (>95 %), and in-house PCR displayed 100 % correct identification of PCR ribotypes 066 and 078. Furthermore, the presence of the PCR enhancer bovine serum albumin (BSA) was found to be related to high sensitivity and low inhibition rate. Rapid laboratory diagnosis of toxigenic C. difficile by RT PCR was accurate

    KW - Clostridium difficile

    KW - Clostridium difficile

    U2 - 10.1007/s10096-014-2284-7

    DO - 10.1007/s10096-014-2284-7

    M3 - Journal article

    VL - 34

    SP - 727

    EP - 736

    JO - European Journal of Clinical Microbiology & Infectious Diseases

    JF - European Journal of Clinical Microbiology & Infectious Diseases

    SN - 0934-9723

    IS - 4

    ER -