Evaluation of available diagnostic methods for Clostridium piliforme in laboratory, rabbits (Oryctolagus cuniculus)
LABORATORY ANIMALS
Authors: Pritt, S.; Henderson, K. S.; Shek, W. R.
Abstract
Recent literature has indicated that animals seropositive for Clostridium piliforme may have negative findings on polymerase chain reaction (IPCR) testing. This study examines and reports on the results of serology, PCR and histopathology tests performed on a group of 20 laboratory rabbits seropositive for C. piliforme using the latest available diagnostic methodologies. The presence of the organism was not confirmed by either IPCR or histopathology in this group of 20 for C. piliforme seropositive rabbits. This presents challenges for laboratory animal veterinarians and scientists wishing to establish the presence of the organism with commonly available diagnostic methods and means that clinical signs of disease in addition to diagnostic results must be interpreted together.
Specific and quantitative detection of PCR products from Clostridium piliforme, Helicobacter bilis, H-hepaticus, and mouse hepatitis virus infected mouse samples using a newly developed electrochemical DNA chip
JOURNAL OF MICROBIOLOGICAL METHODS
Authors: Goto, Kazuo; Horiuchi, Hideki; Shinohara, Haruka; Motegi, Katsumi; Hashimoto, Koji; Hongo, Sadato; Gemma, Nobuhiro; Hayashimoto, Nobuhito; Itoh, Toshio; Takakura, Akira
Abstract
We developed a microfabricated electrochemical DNA chip for detection of polymerase chain reaction (PCR) products from 16S rRNA sequences of Clostridium piliforme (Cp), Helicobacter bilis (Hb) and Helicobacter hepaticus (Hh), and the nucleocapsid protein gene of mouse hepatitis virus (MHV). This chip does not require DNA labeling, and the hybridization signal can be detected as an anodic current. The average anodic currents of 9 (Cp), 5 (Hb), 8 (Hh) and 7 (MHV) PCR positive samples derived from feces of spontaneously infected mice (Cp, Hb and Hh) and MHV-contaminated tumor cells were 27.9 +/- 7.2, 31.9 +/- 8.1, 29.3 +/- 10.1, and 27.6 +/- 3.0 nA, respectively. On the other hand, the average anodic currents of 19 (Cp), 27 (Hb), 18 (Hh), and 13 (MHV) PCR negative samples were 0.3 +/- 2.9, 3.7 +/- 2.4, -1.0 +/- 1.7, and -2.3 +/- 2.7 nA, respectively. The anodic current increased with increasing concentrations of pathogens. For experimentally infected samples, the results of PCR/electrophoresis were in complete accord with those of this system when anodic currents of 6.1 (Cp), 8.5 (Hb), 2.4 (Hh), and 3.1 nA (MHV) were taken as the cut-off value. The results suggested that the electrochemical DNA chip system is useful for specific and quantitative detection of PCR products. (c) 2006 Elsevier B.V. All rights reserved.
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