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Chapter 40 : PCR as a Routine Method for Diagnosis of Legionnaires' Disease

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PCR as a Routine Method for Diagnosis of Legionnaires' Disease, Page 1 of 2

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Abstract:

This chapter evaluates the performance of PCR with respect to sensitivity, specificity, and predictive value of a positive test result with the help of the results obtained by other diagnostic methods used in a routine laboratory. Laboratory results for other diagnostic methods were included if samples were collected within a period of 90 days relative to the sample for PCR. The amplification control thus contained the binding sites of the 16S rDNA primers, was purified by gel electrophoresis, and was added to the mastermix at a concentration producing a distinct amplicon in negative controls without increasing the level of detection for the positive controls. The amplicons of the PCR were analyzed by gel electrophoresis. The authors used two different assays: from 1995 to 1999 an inhouse assay was used and from 1999 the Biotest Urin antigen EIA was used. The performance of PCR as a routine diagnostic method for respiratory infections (Legionnaires' disease) caused by was acceptable, although not all PCR- positive cases could be verified by other diagnostic methods. The sensitivity of PCR is probably higher than any other single method.

Citation: Uldum S, Melbak K. 2002. PCR as a Routine Method for Diagnosis of Legionnaires' Disease, p 213-215. In Marre R, Abu Kwaik Y, Bartlett C, Cianciotto N, Fields B, Frosch M, Hacker J, Lück P (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555817985.ch40

Key Concept Ranking

Legionella pneumophila
0.76875
Legionella micdadei
0.7625
0.76875
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Figures

Image of FIGURE 1
FIGURE 1

Gel electrophoresis of PCR amplicons. Lanes: 1, size marker; 2 and 3, positive samples; 4 and 5, negative samples; 6 and 7, inhibitory samples; 8 and 9, positive controls; 10 and 11, negative controls.

Citation: Uldum S, Melbak K. 2002. PCR as a Routine Method for Diagnosis of Legionnaires' Disease, p 213-215. In Marre R, Abu Kwaik Y, Bartlett C, Cianciotto N, Fields B, Frosch M, Hacker J, Lück P (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555817985.ch40
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References

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1. Povlsen, K.,, J. S. Jensen,, and I. Lind. 1998. Detection of Ureaplasma urealyticum by PCR and biovar determination by liquid hybridization. J. Clin. Microbiol. 36:32113216.
2. Yamamoto, H.,, Y. Hashimoto,, and T. Ezaki. 1993. Comparison of detection methods for Legionella species in environmental water by colony isolation, fluorescent antibody staining, and polymerase chain reaction. Microbiol. Immunol. 37: 617622.

Tables

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TABLE 1

Distribution of the 4,420 patients with samples examined by PCR on positive and negative results

Citation: Uldum S, Melbak K. 2002. PCR as a Routine Method for Diagnosis of Legionnaires' Disease, p 213-215. In Marre R, Abu Kwaik Y, Bartlett C, Cianciotto N, Fields B, Frosch M, Hacker J, Lück P (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555817985.ch40
Generic image for table
TABLE 2

Calculation of the sensitivity, specificity, and predictive value of a positive test result (PVpos.) for PCR from the figures given in Table 1

Citation: Uldum S, Melbak K. 2002. PCR as a Routine Method for Diagnosis of Legionnaires' Disease, p 213-215. In Marre R, Abu Kwaik Y, Bartlett C, Cianciotto N, Fields B, Frosch M, Hacker J, Lück P (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555817985.ch40

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