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Chapter 3 : Detection of Cholera Toxin Genes

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Abstract:

Laboratory diagnosis of cholera has traditionally been based on the phenotypic characteristics of O1, expressed as its morphologic, physiologic, and biochemical properties, including its antigenic composition. Assays based on DNA hybridization, polymerase chain reaction (PCR), and DNA sequencing have been developed for the detection and characterization of the gene. The chapter describes and discusses the advantages and disadvantages of using these techniques in the diagnosis of cholera. Nucleic acid hybridization technology offers distinct advantages over phenotypic identification systems because of its specificity and sensitivity. The format of the hybridization test is important for the practical application of DNA probes for diagnostic purposes. The colony blot system has a great potential for screening a large number of colonies but gives little additional molecular information. A colony blot hybridization assay employing an alkaline phosphatase-labeled 23-bp oligonucleotide probe (CTAP) derived from a specific sequence of was used by Wright and coworkers on a strain collection representing 11 species and toxigenic and nontoxigenic strains of . Nucleic acid-sequencing methods have developed so rapidly over the past 2 decades that comparative sequencing of homologous genes has become almost a standard method in systematic classification of bacteria. The progress in nucleic acid-based methods in recent years has demonstrated that the molecular approach to diagnosis of cholera could very well be the diagnostic choice of the future. Automation and standardization of molecular methods have also increased reproducibility and improved the quality of the laboratory test results.

Citation: Popovic T, Fields P, Olsvik Ø. 1994. Detection of Cholera Toxin Genes, p 41-52. In Wachsmuth I, Blake P, Olsvik Ø (ed), and Cholera. ASM Press, Washington, DC. doi: 10.1128/9781555818364.ch3

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Figures

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Figure 1

CT genes. Numbers designate nucleotide positions, starting with the first nucleotide of the initiation codon ATG of (nucleotide 1) and ending with the third nucleotide of the last codon AAT of (nucleotide 1145).

Citation: Popovic T, Fields P, Olsvik Ø. 1994. Detection of Cholera Toxin Genes, p 41-52. In Wachsmuth I, Blake P, Olsvik Ø (ed), and Cholera. ASM Press, Washington, DC. doi: 10.1128/9781555818364.ch3
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Image of Figure 2
Figure 2

Analysis of PCR products from the genes. Lanes: A, Ol, C9505, human, Mexico, 1992; B, Ol, C9380, human, Mexico, 1992; C, Ol, A1261, human, Bangladesh, 1980; D, Ol, A1263, human, Bangladesh, 1980; E, Ol, C9606, human, Paraguay, 1993; F, Ol, C9607, human, Paraguay, 1993; G, Ol, C9608, human, United States, 1993 H, Ol, C8129, human, Peru, 1992; I, Ol, C8155, human, Peru, 1992; J, C. jejuni, D116, human, United States, 1982; K, Ol, ATCC 14035, human, India, 1949; L, molecular size ladder.

Citation: Popovic T, Fields P, Olsvik Ø. 1994. Detection of Cholera Toxin Genes, p 41-52. In Wachsmuth I, Blake P, Olsvik Ø (ed), and Cholera. ASM Press, Washington, DC. doi: 10.1128/9781555818364.ch3
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References

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Tables

Generic image for table
Table 1

DNA probes for detection of CT genes

Citation: Popovic T, Fields P, Olsvik Ø. 1994. Detection of Cholera Toxin Genes, p 41-52. In Wachsmuth I, Blake P, Olsvik Ø (ed), and Cholera. ASM Press, Washington, DC. doi: 10.1128/9781555818364.ch3
Generic image for table
Table 2

PCR for detection of CT genes

Citation: Popovic T, Fields P, Olsvik Ø. 1994. Detection of Cholera Toxin Genes, p 41-52. In Wachsmuth I, Blake P, Olsvik Ø (ed), and Cholera. ASM Press, Washington, DC. doi: 10.1128/9781555818364.ch3
Generic image for table
Table 3

Sequence differences in B subunits of CT genes

Citation: Popovic T, Fields P, Olsvik Ø. 1994. Detection of Cholera Toxin Genes, p 41-52. In Wachsmuth I, Blake P, Olsvik Ø (ed), and Cholera. ASM Press, Washington, DC. doi: 10.1128/9781555818364.ch3

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