Induction of an electrochemiluminescence sensor for DNA detection of Clostridium perfringens based on rolling circle amplification

Clostridium perfringens is one of the predominant pathogens causing infectious diseases. This work describes the application of a rolling circle amplification (RCA) based electrochemiluminescence sensor for detection of C. perfringens. Firstly, the target DNA is captured by the probes on the pretreated electrode. Subsequently, RCA reaction is executed isothermally. The products of RCA are incubated with hemin, resulting in the decrease of the ECL emission, which is related to the quantity of the target DNA. The ECL-sensor provides the capability of discriminating the target DNA from non-target sequences even with only one base difference, suggesting an advantageous selectivity. Meanwhile, the lowest concentration of the target DNA is 10−15 M, showing satisfactory sensitivity. Therefore, this strategy combines amplification ability of RCA and high sensitivity of ECL, and enables a low fM detection of C. perfringens without the bacterial culture.

[1]  V. Joglekar,et al.  Fatal Clostridium perfringens infection of a liver cyst. , 2003, The Journal of infection.

[2]  Reinhard Niessner,et al.  Automated, high performance, flow-through chemiluminescence microarray for the multiplexed detection of phycotoxins. , 2013, Analytica chimica acta.

[3]  Jianlong Zhao,et al.  A colorimetric method for H1N1 DNA detection using rolling circle amplification. , 2013, The Analyst.

[4]  H. Sakamoto,et al.  Role of alpha-toxin in Clostridium perfringens infection determined by using recombinants of C. perfringens and Bacillus subtilis , 1994, Infection and immunity.

[5]  M. Nicolas-Chanoine,et al.  Typing of Clostridium perfringens strains by use of Random Amplified Polymorphic DNA (RAPD) system in comparison with zymotyping. , 1997, Anaerobe.

[6]  Thomas R Meier,et al.  Gangrenous Clostridium perfringens infection and subsequent wound management in a rhesus macaque (Macaca mulatta). , 2007, Journal of the American Association for Laboratory Animal Science : JAALAS.

[7]  L. Claeys,et al.  Anaerobic cellulitis as the result of Clostridium perfringens: a rare cause of vascular access graft infection. , 2002, Journal of vascular surgery.

[8]  Yang Xiang,et al.  Sensitive and specific HBV genomic DNA detection using RCA-based QCM biosensor , 2013 .

[9]  R. Pearson,et al.  Clostridium perfringens wound infection associated with elastic bandages. , 1980, JAMA.

[10]  V. Chizhikov,et al.  Identification and characterization of Clostridium perfringens using single target DNA microarray chip. , 2004, International journal of food microbiology.

[11]  Shankar Balasubramanian,et al.  Lytic phage as a specific and selective probe for detection of Staphylococcus aureus--A surface plasmon resonance spectroscopic study. , 2007, Biosensors & bioelectronics.

[12]  Ève Bérubé,et al.  Abilities of the mCP Agar Method and CRENAME Alpha Toxin-Specific Real-Time PCR Assay To Detect Clostridium perfringens Spores in Drinking Water , 2013, Applied and Environmental Microbiology.

[13]  D. Arber,et al.  Features of hemolysis due to Clostridium perfringens infection , 2009, International journal of laboratory hematology.

[14]  Su-Hua Huang Gold nanoparticle-based immunochromatographic assay for the detection of Staphylococcus aureus , 2007 .

[15]  Paolo Bertoncello,et al.  Nanostructured materials for electrochemiluminescence (ECL)-based detection methods: recent advances and future perspectives. , 2009, Biosensors & bioelectronics.