[Usage of real time polymerase chain reaction for diagnostics of different tick-borne infections].
OBJECTIVE
To create and test the complex of polymerase chain reaction-based methods for detection of pathogens vectored by ticks in clinical and environmental samples.


METHODS
Real time PCR methods with hybridization-fluorescent detection were developed for detection of tick-borne encephalitis virus, Borrelia burgdorferi sensu lato, Anaplasma phagocytophillum, Erlichia muris/E. chaffeensis, and B. miyamotoi. First four methods were combined in one assay in multiprime format. Efficacy of the assay was assessed by testing of blood samples from patients with tickborreliosis (166 patients), tick-born encephalitis (22 patients) and mixed infection tick-borne encephalitis + borreliosis (21 patients) from Sverdlovsk region.


RESULTS
It was shown that using PCR-based assay for testing the blood samples obtained during admission, it was possible to determine the etiology of disease in 39% of patients, whereas on the basis of serological data diagnosis, as a rule, is made not earlier than on 2nd week of therapy. False-positive results of PCR diagnostics were not observed. Infections caused by Anaplasma or Erlichia were not observed. It was shown that > 50% of cases of tick borreliosis without erythema were caused by B. miyamotoi, whereas B. burgdorferi sensu lato predominated as a causative agent of erythemic form of borreliosis.


CONCLUSIONS
Proposed complex of methods is useful for rapid diagnostics of tick-borne infections including previously unknown infection caused by B. miyamotoi.
Karan'
L S
LS
Koliasnikova
N M
NM
Toporkova
M G
MG
Makhneva
M A
MA
Nadezhdina
M V
MV
Esaulkova
A Iu
AIu
Romanenko
V V
VV
Arumova
E A
EA
Platonov
A E
AE
Maleev
V V
VV
D041081:Anaplasma phagocytophilum / Q000235:genetics / Q000302:isolation & purification; D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics; D001898:Borrelia / Q000235:genetics / Q000302:isolation & purification; D036022:Capsid Proteins / Q000235:genetics; D003937:Diagnosis, Differential; D004536:Ehrlichia / Q000235:genetics / Q000302:isolation & purification; D016873:Ehrlichiosis / Q000175:diagnosis; D004669:Encephalitis Viruses, Tick-Borne / Q000235:genetics / Q000302:isolation & purification; D004675:Encephalitis, Tick-Borne / Q000175:diagnosis; D006801:Humans; D018884:Ixodes / Q000382:microbiology / Q000821:virology; D008193:Lyme Disease / Q000175:diagnosis; D016133:Polymerase Chain Reaction / Q000379:methods*; D012329:RNA, Bacterial / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012680:Sensitivity and Specificity; D017282:Tick-Borne Diseases / Q000175:diagnosis*
D001425:Bacterial Outer Membrane Proteins; D036022:Capsid Proteins; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S; C488283:msp2 protein, Anaplasma phagocytophilum