Comparative evaluation of two Rickettsia typhi-specific quantitative real-time PCRs for research and diagnostic purposes.
Rickettsioses are caused by intracellular bacteria of the family of Rickettsiaceae. Rickettsia (R.) typhi is the causative agent of endemic typhus. The disease occurs worldwide and is one of the most prevalent rickettsioses. Rickettsial diseases, however, are generally underdiagnosed which is mainly due to the lack of sensitive and specific methods. In addition, methods for quantitative detection of the bacteria for research purposes are rare. We established two qPCRs for the detection of R. typhi by amplification of the outer membrane protein B (ompB) and parvulin-type PPIase (prsA) genes. Both qPCRs are specific and exclusively recognize R. typhi but no other rickettsiae including the closest relative, R. prowazekii. The prsA-based qPCR revealed to be much more sensitive than the amplification of ompB and provided highly reproducible results in the detection of R. typhi in organs of infected mice. Furthermore, as a nested PCR the prsA qPCR was applicable for the detection of R. typhi in human blood samples. Collectively, the prsA-based qPCR represents a reliable method for the quantitative detection of R. typhi for research purposes and is a promising candidate for differential diagnosis.
Department of Immunology, Bernhard Nocht Institute for Tropical Medicine, 20359, Hamburg, Germany.
Stefanie
S
Papp
Department of Immunology, Bernhard Nocht Institute for Tropical Medicine, 20359, Hamburg, Germany.
Jessica
J
Rauch
Department of Immunology, Bernhard Nocht Institute for Tropical Medicine, 20359, Hamburg, Germany.
Svenja
S
Kuehl
Department of Immunology, Bernhard Nocht Institute for Tropical Medicine, 20359, Hamburg, Germany.
Ulricke
U
Richardt
Institute for Virology, University Medical Center Gießen and Marburg, 35032, Marburg, Germany.
Christian
C
Keller
Department of Immunology, Bernhard Nocht Institute for Tropical Medicine, 20359, Hamburg, Germany. osterloh@bni-hamburg.de.
Anke
http://orcid.org/0000-0002-4078-1070
A
Osterloh
D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics; D001769:Blood / Q000382:microbiology; D006801:Humans; D008074:Lipoproteins / Q000235:genetics; D008565:Membrane Proteins / Q000235:genetics; D016513:Mice, SCID; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012286:Rickettsia typhi / Q000235:genetics / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D014437:Typhus, Endemic Flea-Borne / Q000175:diagnosis* / Q000382:microbiology
D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D008074:Lipoproteins; D008565:Membrane Proteins; C082160:prsA protein, bacteria
Differential diagnosis; Endemic typhus; Quantification; R. typhi; qPCR
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