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Chapter 33 : Clinical Validation of Diagnosis of Infections

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Clinical Validation of Diagnosis of Infections, Page 1 of 2

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Abstract:

is the etiologic agent of Legionnaires' disease, which is a very serious illness with pneumonia and a mortality of 15 to 20%. In addition to , more than 40 other spp. are known (designated in this chapter as non- species), of which several species have been shown to be pathogenic for humans. To improve diagnosis of infections, the authors designed a novel PCR to specifically amplify all DNA. Specific probes LPN and LSPP were used for discrimination of, respectively, and non- species. The sensitivity of PCR was determined by spiking bacteria in negative clinical material. The lower detection limit was found to be at least 0.1 CFU. A total of 208 samples from 208 patients clinically suspected of legionellosis were subjected to the PCR/probe procedure. Comparison of results of PCR, serology, culture, and urine antigen detection revealed that PCR-based detection of , and serology in reconvalescent serum, yielded 2.8-fold more positives than culture. PCR may provide an important contribution to an early diagnosis of legionellosis. PCR is the only method suitable to diagnose non- infections. Future studies could include sequencing of non- PCR products to get an impression of the distribution of species in patient samples.

Citation: van der Zee A, Verbakel H, de Jong C, Pot R, Peeters M, Bergmans A, Schellekens J. 2002. Clinical Validation of Diagnosis of Infections, p 189-192. 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.ch33

Key Concept Ranking

Legionella pneumophila
0.67142856
Pseudomonas aeruginosa
0.54285717
0.67142856
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Figures

Image of FIGURE 1
FIGURE 1

Schematic presentation of the PCR/probe procedure. LEG1: 5′ -TACCTACCCTTGACATACAGTG-3′, LEG2: 5′-CTTCCTCCGGTTTGTCAC-3′ , LPN-B: 5′-ATGTGATGGTGGGGACTCT 3′, LSPP-B:5′ - CGTAACGAGCGCAACCC-3′ .

Citation: van der Zee A, Verbakel H, de Jong C, Pot R, Peeters M, Bergmans A, Schellekens J. 2002. Clinical Validation of Diagnosis of Infections, p 189-192. 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.ch33
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Image of FIGURE 2
FIGURE 2

(A) The sensitivity of PCR detection of non- Twofold dilutions of bacterial cells were used in PCR after processing in addition to internal control. Lanes 1 to 8 correspond to a range of 10 to .08 cells per reaction. The sizes of amplified fragments are indicated on the right. Below, PCR products are hybridized to probe LPN-B. (B) The specificity of the PCR detection of non- PCR was performed on the equivalent of 100 cells of two different clinical isolates of (lanes 1, 2), (lane 3), (lane 4), (lane 5), (lane 6), (lane 7), (lane 8), (lane 9), (lane 10), (lane 11), and (lane 12). Below, PCR products are hybridized to probe LSPP-B at 60°C, and to probe LPN-B at 61°C.

Citation: van der Zee A, Verbakel H, de Jong C, Pot R, Peeters M, Bergmans A, Schellekens J. 2002. Clinical Validation of Diagnosis of Infections, p 189-192. 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.ch33
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References

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1. Ching, W. T. W.,, and R. D. Meyer. 1987. Legionella infections. Infect. Dis. Clin. N. Am. 1: 595614.
2. Harrison, T. G.,, and A. G. Taylor,. 1988. The diagnosis of legionnaires’ disease by estimation of antibody levels, p. 113136. In T. G. Harrison, and A. G. Taylor (ed.), A Laboratory Manual for Legionella. John Wiley & Sons Ltd., New York, N.Y..
3. Marston, B.,, J. F. Plouffe,, R. F. Breiman,, T. M. File, Jr.,, R. F. Benson,, M. Moyenud-den,, W. L. Thacker,, K.-H. Wong,, S. Skelton,, B. Hackman,, S. J. Salstrom,, J. M. Barbaree,, and the Community-Based Pneumonia Incidence Study Group. 1993. Preliminary findings of a community-based pneumonia incidence study, p. 3637. In J. M. Barbaree,, R. F. Breiman,, and A. P. Dufour (ed.), Legionella: Current Status and Emerging Perspectives. American Society for Microbiology, Washington, D.C..
4. Plouflfe, J. F.,, T. M. File,, R. F. Breiman,, B. A. Hackman,, S. J. Salstrom,, B. J. Marston,, B. S. Fields, and the Community-Based Pneumonia Incidence Study Group. 1995. Reevaluation of the definition of Legionaires disease: use of the urinary antigen assay. Clin. Infect. Dis. 20:12861291.

Tables

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TABLE 1

Relative sensitivities and specificities of diagnostic methods

Citation: van der Zee A, Verbakel H, de Jong C, Pot R, Peeters M, Bergmans A, Schellekens J. 2002. Clinical Validation of Diagnosis of Infections, p 189-192. 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.ch33

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