Evaluation of real-time PCR targeting the lipL32 gene for diagnosis of Leptospira infection
Autor: | Mateja Logar, Daša Podgoršek, Eva Ružić-Sabljić, Emil Pal, Tjaša Cerar, Zvonko Baklan, Andrea Pavlović, Tatjana Remec |
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Rok vydání: | 2019 |
Předmět: |
Microbiology (medical)
Lipl32 Lipoproteins lcsh:QR1-502 Urine Biology Real-Time Polymerase Chain Reaction Microbiology Sensitivity and Specificity lcsh:Microbiology Serology 03 medical and health sciences Leptospira Agglutination Tests medicine Humans Leptospirosis 030304 developmental biology 0303 health sciences Bacteriological Techniques 030306 microbiology Sequence Analysis DNA medicine.disease biology.organism_classification Virology Agglutination (biology) Real-time polymerase chain reaction Blood Early Diagnosis PCR Parasitology Molecular Diagnostic Techniques biology.protein Antibody Bacterial Outer Membrane Proteins Research Article |
Zdroj: | BMC Microbiology BMC Microbiology, Vol 20, Iss 1, Pp 1-9 (2020) |
ISSN: | 1471-2180 |
Popis: | Background Different diagnostic methods have been used for the laboratory confirmation of leptospirosis. Molecular diagnostic techniques are not only faster and more sensitive than culture analysis, but can also detect a Leptospira infection before the appearance of antibodies. The aim of the present study was to analyze and compare two different PCR approaches applied to blood and urine specimens obtained from patients with clinical manifestations that were suggestive of leptospirosis. Furthermore, the results of these different PCR approaches were compared with the results of culture and serology analyses. Results A total of 400 samples (234 blood or 58.5% and 166 urine of 41.5%) from 310 Slovenian patients with clinical manifestations suggestive of leptospirosis were tested using conventional PCR assays targeting the rrs gene and RT-PCR targeting the lipL32 gene. Additionally, culture, serology and sequence analysis were performed for the majority of these samples. The PCR and RT-PCR results were concordant in 376 out of 400 of these samples (94.0%). Conventional PCR was positive for 27 out of 400 samples (6.8%) and RT-PCR was positive for 47 out of 400 samples (11.8%). Culture and microscopic agglutination tests supported these diagnoses. Conclusions A comparison of the two PCR methods indicated that the RT-PCR targeting of the lipL32 gene was faster, more sensitive and more specific for the determination of Leptospira DNA in these clinical samples. |
Databáze: | OpenAIRE |
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