DETECTION OF LYME-BORRELIOSIS, BARTONELLIOSIS AND EPSTEIN-BARR VIRAL INFECTION IN PATIENTS WITH LYMPHADENOPATHY, RESIDENTS OF TERNOPIL REGION

Abstract

SUMMARY. The aim – to examine residents of the Ternopil region with lymphadenopathy using serological and molecular genetic methods for the presence of Lyme-borreliosis, bartonellosis, EBV infection and their combination.

Patients and methods. In the Center for Lyme-borreliosis and other tick-borne diseases research at the I. Horbachevsky Ternopil National Medical University were examined 36 patients aged 19 to 76 who were diagnosed with lymphadenopathy (LAP). There were 9 (25.0 %) men, 27 (75.0 %) women. 26 (72.2 %) persons lived in the city, 10 (27.8 %) lived in the countryside.

For detection of IgM and/or IgG to B. burgdorferi s. l. in blood serum, a two-stage scheme (ELISA and immunoblot) was used, using the test systems of the company Euroimmun AG (Germany). The obtained results were analyzed in accordance with the recommendations of the manufacturer of the test systems. Antibodies of class G to Bartonella henselae and Bartonella quintana in the blood serum of patients were determined using the method of multiplex indirect immunofluorescence, using the test systems «Mosaic for Bartonella henselae/Bartonella quintana (IgG)», the company Euroimmun AG (Germany), BIOCHIP technology, which contained fluorescein-labeled antigens of the specified Bartonella species.

The results of the determination of specific antibodies to B. henselae and B. quintana and the indicated antigens of EBV-infection were evaluated in the field of view of a fluorescent microscope (Olympus IX70, glasses×10, lens×20;40) by the bright green glow of the antigen-antibody immune complex labeled with fluorescein.

For the diagnosis of EBV infection, the indirect immunofluorescence multiplex reaction (BIOCHIP technology) was used. The test system “BIOCHIP Sequence EBV (with avidity determination)” (EUROIMMUN, Germany) was used, which contains the capsid antigen and its proteins gp125 and p19, nuclear and early antigens of EBV.

To establish the active or latent phase of chronic EBV infection, real-time PCR was used, with the help of which EBV DNA was determined in the blood plasma and saliva of the examined patients. The active phase of chronic EBV-infection was diagnosed by the presence of EBV DNA in blood and saliva (in both or one sample; detection range – 10³-10⁷ copies/ml). In the absence of viral DNA in blood or saliva, the latent phase of chronic EBV infection was established.

Results. 50.0 % of patients with lymphadenopathy were bitten by ticks, mostly in the lower limbs, significantly more often during their stay at the forest and gardens, p<0.05.

The alternate use of ELISA and immunoblot made it possible to diagnose Lyme borreliosis in 55.6 % of patients with lymphadenopathy. Specific IgG antibodies only to B. henselae were diagnosed in the blood serum of 13.9 % of patients with lymphadenopathy associated with Lyme borreliosis. The indirect immunofluorescence multiplex reaction method (BIOCHIP technology) made it possible to diagnose chronic EBV infection in all patients with lymphadenopathy. With the help of polymerase chain reaction in real time, the active phase of chronic EBV-infection was established in 52.8 % of patients, the latent phase – in 47.2 % of patients. 72.2 % of patients with lymphadenopathy were diagnosed with Lyme borreliosis, bartonellosis caused by B. henselae, and the active phase of chronic EBV-infection both as individually and so in various combinations.

Conclusion. Patients with lymphadenopathy should be screened for possible concurrent Lyme disease, bartonellosis, and chronic EBV-infection. The indirect immunofluorescence multiplex reaction method (BIOCHIP technology) with determination of eight specific antibodies at the same time for the diagnosis of EBV-infection in patients with lymphadenopathy, residents of the Ternopil region, was used for the first time and demonstrated high informativeness.