The Use of Multiplex Phosphorescence Analysis (PHOSPHAN<sup>TM</sup>) and Polymerase Chain Reaction for Laboratory Diagnosis of Ixodid Tick-borne Borrelioses

Abstract

In this report, we evaluated the performance of C6 peptide based multiplex Phosphorescence Analysis (PHOSPHANTM) and Polymerase Chain Reaction (nested PCR) for laboratory diagnosis of Ixodid Tick-borne Borrelioses (ITBB). The study was conducted on 155 patients with localized and disseminated stages of the disease, the cases of mixed infection with ITBB and human granulocytic anaplasmosis including. Positive PHOSPHAN reactions were observed in 78 ± 7.7% of patients with erythema migrans (EM) and 91 ± 11.7% of patients without cutaneous manifestations of the disease. The frequency of PCR positive samples was lower, 26 ± 8.2% and 72 ± 17.1% respectively. The maximum frequency of positive samples detected by both methods was mainly observed at 2 - 4 week from the onset of the disease (or 22 - 35 day after tick bite). In general, PHOSPHAN provided serologic confirmation of the disease in 52 of 55 (94.5 ± 6.2%) patients, whose blood contained Borrelia DNA. Only 3 patients tested positive in PCR (1 - with EM and 2 - without this skin manifestation) were seronegative. These data confirmed the high efficiency of PHOSPHAN method for serologic verification of ITBB both at localized and disseminated stages of the disease. The use of PCR (in addition to PHOSPHAN) is appropriate within a certain period of time (no later than 2 - 3 weeks from the onset of the disease) to clarify the diagnosis in seronegative patients having clinical signs of disseminated non-cutaneous form of ITBB, or atypical cutaneous manifestations of erythematous form of the disease.