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A quantitative polymerase chain reaction method for the detection in avian faeces of salmonellas carrying the spvR gene

Published online by Cambridge University Press:  15 May 2009

J. Mahon  and
A. J. Lax
J. Mahon
Affiliation:
AFRC Institute for Animal Health, Compton Laboratory, Compton, Newbury, Berkshire, RG16 0NN, UK
A. J. Lax
Affiliation:
AFRC Institute for Animal Health, Compton Laboratory, Compton, Newbury, Berkshire, RG16 0NN, UK
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A quick, semi-quantitative method of detecting Salmonella species which contain the virulence plasmid has been developed using the polymerase chain reaction (PCR). A pair of primers have been synthesized encompassing a 500 bp fragment of the spvR virulence gene. Competitor DNA consisting of the spvR gene with a 94 bp deletion situated between the primer recognition sequences, was cloned into a plasmid vector. Co-amplification of the ‘unknown’ target salmonella DNA with known quantities of competitor DNA in the same reaction tube gave PCR products of 500 and 406 bp respectively. Visual assessment of the ratio of the two products on ethidium bromide stained agarose gels provided an estimate of the approximate number of salmonella cells present in avian faeces.

The technique could be applied to detect quantifiably any non-host DNA in clinical samples if a suitable DNA sequence for primer construction is available.

Type
Research Article
Information
Epidemiology & Infection , Volume 111 , Issue 3 , December 1993 , pp. 455 - 464
Copyright
Copyright © Cambridge University Press 1993

References

1.Methods for the isolation and identification of Salmonellae (other than Salmonellae typhi) from water and associated materials. London: HMSO, 1982.Google Scholar
2.Widjojoatmodjo, MN, Fluit, AC, Torensma, R, Keller, BHI, Verhoef, J. Evaluation of the magnetic immuno PCR assay for rapid detection of Salmonella. Eur J Clin Micro Infect Dis 1991; 10: 935–8.CrossRefGoogle ScholarPubMed
3.Wernars, K, Heurelman, P, Chakreboty, T, Notermans, SHW. Use of the polymerase chain reaction for direct detection of Listeria monocytogenes in soft cheese. J Appl Bact 1991; 70: 121–6.CrossRefGoogle ScholarPubMed
4.Wernars, K, Delfgou, E, Soentoro, PS, Notermans, SHW. Successful approach for detection of low numbers of entertoxigenic Escherichia coli in minced meat by using the polymerase chain reaction. Appl Env Micro 1991; 57: 1914–19.CrossRefGoogle Scholar
5.Bej, AK, Steffan, RJ, DiCesare, J, Haff, L, Atlas, RM. Detection of coliform bacteria in water by the polymerase chain reaction and gene probes. Appl Env Micro 1990; 56: 307–14.CrossRefGoogle ScholarPubMed
6.Wang, R-F, Cao, WW, Johnson, MG. Development of cell surface protein associated gene probe specific for Listeria monocytogenes and detection of the bacteria in food by PCR. Mol Cell Probes 1992; 6: 119–29.CrossRefGoogle ScholarPubMed
7.Tsai, Y-L, Olsen, BH. Detection of low numbers of bacterial cells in soils and sediments by polymerase chain reaction. Appl Env Micro 1992; 58: 754–7.CrossRefGoogle ScholarPubMed
8.Frankel, G, Riley, L, Giron, A, et al. Detection of Shigella in faeces using DNA amplification. J Infect Dis 1990; 161: 1252–6.CrossRefGoogle ScholarPubMed
9.Allard, A, Girones, R, Juto, P. Wadell, G. Polymerase chain reaction for detection of adenoviruses in stool samples. J Clin Micro 1990; 28: 2659–67.CrossRefGoogle ScholarPubMed
10.Davis, GR, Blumayer, K, DiMichele, LJ, et al. Detection of human immunodeficiency virus type I in AIDS patients using amplification-mediated hybridization analysis: reproducibility and quantitative limitations. J Infect Dis 1990; 162: 13–20.CrossRefGoogle ScholarPubMed
11.Steiger, M, Demolliere, C, Ahlborn-laake, L, Mous, J. Competitive polymerase chain reaction assay for quantitation of HIV-1 DNA and RNA. J Virol Meth 1991; 34: 149–60.CrossRefGoogle Scholar
12.Holodiniy, M, Katzenstein, DA, Sengupta, S, et al. Detection and quantification of human immunodeficiency virus RNA in patient serum by use of the polymerase chain reaction. J Infect Dis 1991; 163: 862–6.CrossRefGoogle Scholar
13.Wang, AM, Doyle, MV, Mark, DF. Quantitation of mRNA by the polymerase chain reaction. Proc Natl Acad Sci USA 1989; 86: 9717–21.CrossRefGoogle ScholarPubMed
14.Seibert, PD, Larrick, JW. Competitive PCR. Nature 1992; 359: 557–8.CrossRefGoogle Scholar
15.Becker-Andre, M, Hohlbrock, K. Absolute mRNA quantification using the polymerase chain reaction (PCR). A novel approach by a PCR aided transcript titration assay (PATTY). Nucleic Acids Res 1989; 17: 9437–46.CrossRefGoogle ScholarPubMed
16.Gilliland, G, Perrin, S, Blanchard, K, Bunn, HF. Analysis of cytokine mRNA and DNA: detection and quantification by competitive polymerase chain reaction. Proc Natl Acad Sci USA 1990; 87: 2725–9.CrossRefGoogle Scholar
17.Nedelman, J, Heagerly, P, Lawrence, C. Quantitative PCR: procedures and precisions. Bull Math Biol 1992; 54: 477–502.CrossRefGoogle Scholar
18. yyStatutory Instruments. The Poultry Breeding Flocks and Hatcheries (Registration and Testing) Order No. 1963. London: HMSO, 1989.Google Scholar
19.Miles, AA, Misra, SS, Irwin, JO. The estimation of the bactericidal power of the blood. J Hyg 1938; 38: 732–49.Google ScholarPubMed
20.Pullinger, GD, Baird, GD, Williamson, CM, Lax, AJ. Nucleotide sequence of a plasmid gene involved in the virulence of salmonellas. Nucleic Acids Res 1989; 17: 7983.CrossRefGoogle ScholarPubMed
21.Sambrook, J, Fritsch, EF, Maniatis, T. Molecular cloning: a laboratory manual. Cold Spring Harbor: Cold Spring Harbor Laboratory Press, NY 1989.Google Scholar
22.Williamson, CM, Baird, GD, Manning, EJ. A common virulence region on plasmids from eleven serotypes of Salmonella. J Gen Micro 1988; 134: 975–82.Google ScholarPubMed
23.Tinge, SA, Curtiss, R. Isolation of the replication and partitioning region of the Salmonella typhimurium virulence plasmid and stabilization of heterologous replicons. J Bact 1990: 172: 5266–77.CrossRefGoogle ScholarPubMed
24.Gulig, PA, Danbara, H, Guiney, DG, Lax, AJ, Norel, F, Rhen, M. Molecular analysis of spr virulence genes of the salmonella virulence plasmids. Mol Micro 1993: 7: 825–30.CrossRefGoogle Scholar
25.Van Pouske, LSG. Salmonella-TEK, a rapid screening method for Salmonella species in food. Appl Env Micro 1990; 56: 924–7.CrossRefGoogle Scholar
26.Choi, D, Tsang, RSW, Ng, MH. Sandwich capture ELISA by a murine monoclonal antibody against a genus-specific LPS epitope for the detection of different common serotypes of salmonellas. J Appl Bact 1992; 72: 134–8.CrossRefGoogle ScholarPubMed
27.Aguirre, PM, Cacho, JB, Folgueira, L, Lopez, M. Garcia, J, Velasco, AC. Rapid fluorescence method for screening Salmonella species from enteric differential agars. J Clin Micro 1990: 28: 148–9.CrossRefGoogle ScholarPubMed
28.Ruiz, J, Sempere, MA, Valera, MC, Gomez, P. Modification of the methodology of stool culture for Salmonella detection. J Clin Micro 1992; 30: 525–6.CrossRefGoogle ScholarPubMed
29.Parmar, N, Easter, MCR, Forsythe, SJ. The detection of S. enteritidis and S. typhimurium using immunomagnetic separation and conductance microbiology. Lett Appl Micro 1992: 15: 175–8.CrossRefGoogle ScholarPubMed
30.Gibson, DM, Coombs, P, Pimbley, DW. Automated conductance methods for detection of Salmonella in foods; collaborative study. J AOAC International 1992; 75: 293–302.CrossRefGoogle Scholar
31.Manafi, M, Sommer, R. Comparison of 3 rapid screening methods for Salmonella species: MUCAP test, MicroscreenR Latex and Rambach agar. Lett Appl Micro 1992: 15: 175–8.Google Scholar
32.Widjojoatmodjo, MN, Fluit, AC, Torensma, R, Verdonk, GPHT, Verhoef, J. The magnetic immuno polymerase chain reaction assay for direct detection of Salmonella in faecal samples. J Clin Micro 1992; 30: 3195–9.CrossRefGoogle Scholar
33.Ursi, JP, Ursi, D, Leven, M, Pattyn, S. Utility of an internal control for the polymerase chain reaction. APMIS 1992; 100: 635–9.CrossRefGoogle ScholarPubMed
34.Khan, G, Kangro, HO, Coates, PJ. Health, RB. Inhibitory effects of urine on the polymerase chain reaction for cytomegalovirus DNA. J Clin Pathol 1992; 44: 360–5.CrossRefGoogle Scholar