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Chapter 12 : Detection of DNA in Serum Samples from Patients with Legionnaires’ Disease

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Detection of DNA in Serum Samples from Patients with Legionnaires’ Disease, Page 1 of 2

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Abstract:

Diagnosis of Legionnaires’ disease (LD) in patients with pneumonia is based on phenotypic (culture, serologic testing, antigen detection in urine) and genotypic PCR methods. This chapter assesses the sensitivity and specificity of DNA detection using PCR on serum samples from patients with proven LD and patients with infections other than . Serum samples from two healthy volunteers were included as negative controls after every four samples. One patient was admitted to the emergency department of our hospital with proven LD caused by serogroup 1 (urinary antigen test positive, culture positive, and sputum PCR positive). From this patient, consecutive serum samples were collected for -specific, real-time PCR. The results of PCR showed an increase in Ct values (corresponding with a logarithmic decrease of bacterial DNA) in the course of time and were found to mirror the clinical condition of the patient and c-reactive protein values during the acute stage of infection. Serologic methods to diagnose infections are highly sensitive, but their utility is generally limited to epidemiologic studies due to the time lag needed to detect seroconversion. The advantages of PCR for the detection of DNA in serum are evident; serum samples are readily obtainable and can be processed within a working day.

Citation: Diederen B, de Jong C, Marmouk F, Kluytmans J, Peeters M, van der Zee A. 2006. Detection of DNA in Serum Samples from Patients with Legionnaires’ Disease, p 47-50. In Cianciotto N, Kwaik Y, Edelstein P, Fields B, Geary D, Harrison T, Joseph C, Ratcliff R, Stout J, Swanson M (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555815660.ch12

Key Concept Ranking

Real-Time PCR
0.7691922
Legionella pneumophila
0.6321857
0.7691922
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Figures

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FIGURE 1

Results of real-time PCR on serum from a patient with proven LD requiring intensive care unit admission. An increase in Ct value corresponds with a decrease in bacterial DNA.

Citation: Diederen B, de Jong C, Marmouk F, Kluytmans J, Peeters M, van der Zee A. 2006. Detection of DNA in Serum Samples from Patients with Legionnaires’ Disease, p 47-50. In Cianciotto N, Kwaik Y, Edelstein P, Fields B, Geary D, Harrison T, Joseph C, Ratcliff R, Stout J, Swanson M (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555815660.ch12
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References

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1. Fields, B. S.,, R. F. Benson, and, R. E. Besser. 2002. Legionella and Legionnaires’ disease: 25 years of investigation. Clin. Microbiol. Rev. 15:506526.
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10. Yu, V. L.,, J. F. Plouffe,, M. C. Pastoris,, J. E. Stout,, M. Schousboe,, A. Widmer,, J. Summersgill,, T. File,, C. M. Heath,, D. L. Paterson, and, A. Chereshsky. 2002. Distribution of Legionella species and serogroups isolated by culture in patients with sporadic community-acquired Legionellosis: an international collaborative study. J. Infect. Dis. 186:127128.

Tables

Generic image for table
TABLE 1

16S rRNA, 5S rRNA, and primer and probe sequences used in PCR (bp; basepairs)

Citation: Diederen B, de Jong C, Marmouk F, Kluytmans J, Peeters M, van der Zee A. 2006. Detection of DNA in Serum Samples from Patients with Legionnaires’ Disease, p 47-50. In Cianciotto N, Kwaik Y, Edelstein P, Fields B, Geary D, Harrison T, Joseph C, Ratcliff R, Stout J, Swanson M (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555815660.ch12

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