Development of the ELISA Test System for Quantitative Determination of IgG to the Varizella zoster virus in Human Serum and Assessment of its Diagnostic Efficiency

Relevance. The introduction of Varicella vaccine prophylaxis explains the need to develop a methodology for monitoring the vaccination effectiveness and the intensity of population immunity. This problem can be solved using quantitative immunoassay methods. Aim. Development of an enzyme-linked immunosorbent assay for the concentration of class G immunoglobulins (AB) to Varicella zoster virus (VZV) determining and assessing its functional characteristics and diagnostic efficiency. Materials and methods. Recombinant antigen GE VZV. WHO International Standard for Antibodies to VZV W1044. Blood serum samples from healthy people and patients with Chickenpox and Herpes zoster, blood serum samples containing IgG antibodies to herpes simplex viruses of the first and second types, cytomegalovirus, Epstein-Barr virus. Anti-VZV ELISA (IgG) reagent kit (Euroimmun, Germany). Indirect enzyme-linked immunosorbent assay. Immunization of animals with recombinant antigen GE, isolation, and purification of specific antibodies. Conjugation of monoclonal antibodies to human IgG with antibodies to antigen GE and with horseradish peroxidase. Results. An enzyme-linked immunosorbent assay in «an indirect» format has been developed to determine the specific antibodies to VZV concentration (IU/ml) in human serum/plasma. An artificial calibrator for determining the concentration of AB-VZV had been synthesized and standardized according to the International WHO-standard W1044. The main functional characteristics of the developed enzyme-linked immunosorbent assay are determined in accordance with GOST 51352-2013. The diagnostic kit was tested on blood serum samples from children with chickenpox (n = 43), adults with Herpes zoster (n = 158), healthy individuals (n = 781). The diagnostic sensitivity of the test system was 85%, the diagnostic specificity was 87% according to the ROC analysis. The absence of cross-reactivity of the test system was shown on samples with serological markers of other herpesvirus infections (n = 94). Comparative trials of the developed test system and its commercial analog, the Anti-VZV ELISA (IgG) reagent kit, did not reveal statistically significant differences between their functional characteristics. Conclusions. The developed test system for determining of the AB-VZV concentration in human serum/plasma in terms of its functional characteristics meets the GOST requirements, is characterized by high diagnostic efficiency, can be used to monitor the effectiveness of vaccine prophylaxis and strength of population immunity, as well as to assess the immune response in chickenpox and Herpes zoster.

Antalya İli Kumluca İlçesindeki Biber Yetiştirilen Seralarda Domates Lekeli Solgunluk Virüsü (Tomato spotted wilt tospovirus-TSWV)’ nün Yaygınlığı ve Konukçu Yabancı Ot Türlerinin Belirlenmesi

There are many viruses that infect pepper and limit its production. Among these viruses, Tomato spotted wilt tospovirus (TSWV) infects crops in 35 plant families that are economically important, including pepper. In the present study; totaly 156 leaf samples were collected, including 57 from pepper plants showing virus-like symptoms and 99 from weeds and/or plants other than peppers in and around the greenhouse, through surveys carried out in pepper greenhouses in Kumluca district of Antalya province, from September to December 2020. Then, the plant leaf samples were tested to determine TSWV infections by the Double Antibody Sandwich Enzyme-linked Immunosorbent Assay (DAS-ELISA) method. According to result of the tests, it was determined that 55.76% of the tested leaf samples were infected with TSWV, while this rate was determined as 96.49% for pepper samples and 32.32% for other plant samples. During the survey studies, it was revealed that the leaf samples of 13 out of 31 weed and different plant species except pepper were infected with at least one of the viruses. In addition, pepper plants showing symptoms TSWV-like symptoms in pepper greenhouses were counted during the survey, and the prevalence of this virus disease was calculated on the basis of Kumluca district and neighborhoods. As a result of these calculations, the prevalences of TSWV; for Kumluca, Mavikent, Beykonak, Salur, Hacıveliler, Adrasan, Merkez, and Kavakköy were determined as 26.93%, 26.92%, 32.27%, 20.66%, 21.13%, 17.66%, 13%, and 25%, respectively.

Imunoenzimske metode u procjeni kvalitete meda i pčelinjih proizvoda

Imunoenzimska ELISA metoda (engl. Enzyme-Linked ImmunoSorbent Assay) se istražuje u posljednje vrijeme kao brza, jednostavna i visokospecifična analitička metoda za unaprjeđenje i standardizaciju kvalitete meda i pčelinjih proizvoda. Med i pčelinji proizvodi se osim zbog nutritivne vrijednosti učestalo koriste i zbog brojnih pozitivnih bioloških učinaka, a ELISA metoda svoju primjenu u standardizaciji njihove kvalitete može pronaći kod utvrđivanja autentičnosti, botaničkog podrijetla te razlikovanja uzoraka prirodnog meda. Metoda se razvija u svrhu određivanja pčelinjih proteina i peptida, kao što su apalbumin-1 i defensin-1, a koji mogu predstavljati biljege izvornosti. Imunoenzimska ELISA metoda primjenu pronalazi i u određivanju peludi određene biljne vrste u svrhu utvrđivanja botaničkog podrijetla, te pri procjeni kvalitete određivanjem enzima, kao i u svrhu razlikovanja uzoraka prirodnog meda određivanjem specifičnih proteina.