scholarly journals STUDY OF ELISA TEST-SYSTEMS OF DIFFERENT FORMATS FOR DETECTION OF MEASLES VIRUS SPECIFIC IgM IN DIFFERENT GEOGRAPHIC ZONES

2018 ◽  
Vol 8 (2) ◽  
pp. 230-234
Author(s):  
M. A. Bichurina ◽  
N. V. Zheleznova ◽  
I. N. Lavrentieva ◽  
A. Yu. Antipova ◽  
L. B. Kulyashova ◽  
...  

Detection of the measles virus (MV) specific IgM antibodies in blood serum of patients is considered to be the main standard for the  laboratory confirmation of measles diagnosis, the test being  acknowledged by WHO. As it was demonstrated earlier the specific  IgM antibodies as the marker of the acute MV infection were  detected in 97.2–100% of blood serum samples from patients using  the ELISA test-systems of the “capture” format (Microimmun Ltd.  and Vector Best). In case when the ELISA test-system of the  “indirect” format (Siemens, Germany) was used only 63.9% of these sera turned to be IgM positive. And on the contrary using the  “indirect” format ELISA test-system Euroimmun, Germany, for  detection of the MV specific IgM the false positive results were  obtained.The aim of the study was the comparative evaluation of  the different format ELISA test-systems used for the detection of the  MV specific IgM antibodies in blood sera of patients and healthy  adults collected in different geographic zones.Materials and  methods. In total 108 serum specimens collected in 2015–2017 were studied: from healthy adult Guineans, residents of the Republic of Guinea (RG); patients aged 1–70 with the initial  “infectious mononucleosis”, “infectious cytomegalovirus” and  “rubella” diagnosis and taken from the bank of sera in the  Subnational Measles/Rubella laboratory, StP Measles/Rubella RC in  NWFR. The MV specific IgM antibodies were detected using the commercial ELISA test-systems “VectoMeasles-IgM” (Vector-Best,  Russia) (“capture” format) and “Anti-Measles Virus ELISA IgM (NP)”  (Euroimmun Medizinische Labordiagnostik AG, Germany) («indirect»  format). The specific Epshtein-Barr Virus (EBV) IgM and IgG  antibodies were detected with the commercial ELISA test-systems «DS-ELISA-anti-EBV-VCA-M», «DS-ELISA-anti-EBV-EA-G» and «DS-ELISA-anti-EBV-NA-G» (“Diagnostic Systems”, Russia).Results and discussion. The MV specific IgM antibodies were not revealed in the total of 108 blood serum samples from the healthy adults and patients, residents of the Russia and of the RG, with the “capture”  format “VectoMeasles-IgM” ELISA test-system. The absence of the  acute MV infection was also confirmed by the high measles immunity  level (i.e. IgG MV antibodies titers) as well as by detection of the IgG antibodies of high avidity. At the same time in 6  of 108 total sera (5.5%) IgM MV antibodies were detected with the «indirect» format ELISA test system Euroimmun, Germany. In these 6 sera the EBV specific antibodies were also evidenced. The results  obtained demonstrate the nonspecific reaction due to the possible  reactivity with anti-EBV antibodies. Besides this the different  percentage of the false positive reactions in sera from healthy  adults, residents of the RG and residents of Russia was determined  — 8.5±4.0% and 3.2±2.2% correspondently. Thus the preliminary  results, and to get the final results for general conclusions increase  of the total amount of the clinical specimens under studying is of extremely importance.

2021 ◽  
Vol 15 (3) ◽  
pp. e0009280
Author(s):  
Petra Emmerich ◽  
Ronald von Possel ◽  
Christina Deschermeier ◽  
Salih Ahmeti ◽  
Lindita Berisha ◽  
...  

Crimean-Congo Hemorrhagic Fever Virus (CCHFV) is a geographically widespread tick-borne arbovirus that has been recognized by the WHO as an emerging pathogen needing urgent attention to ensure preparedness for potential outbreaks. Therefore, availability of accurate diagnostic tools for identification of acute cases is necessary. A panel comprising 121 sequential serum samples collected during acute, convalescent and subsided phase of PCR-proven CCHFV infection from 16 Kosovar patients was used to assess sensitivity. Serum samples from 60 healthy Kosovar blood donors were used to assess specificity. All samples were tested with two IgM/IgG immunofluorescence assays (IFA) from BNITM, the CCHFV Mosaic 2 IgG and IgM indirect immunofluorescence tests (IIFT) from EUROIMMUN, two BlackBox ELISAs for the detection of CCHFV-specific IgM and IgG antibodies (BNITM), two Anti-CCHFV ELISAs IgM and IgG from EUROIMMUN using recombinant structural proteins of CCHFV antigens, and two ELISAs from Vector-Best (IgM: μ-capture ELISA, IgG: indirect ELISA using immobilized CCHFV antigen). Diagnostic performances were compared between methods using sensitivity, specificity, concordance and degree of agreement with particular focus on the phase of the infection. In early and convalescent phases of infection, the sensitivities for detecting specific IgG antibodies differed for the ELISA test. The BlackBox IgG ELISA yielded the highest, followed by the EUROIMMUN IgG ELISA and finally the VectorBest IgG ELISA with the lowest sensitivities. In the subsided phase, the VectorBest IgM ELISA detected a high rate of samples that were positive for anti-CCHFV IgM antibodies. Both test systems based on immunofluorescence showed an identical sensitivity for detection of anti-CCHFV IgM antibodies in acute and convalescent phases of infection. Available serological test systems detect anti-CCHFV IgM and IgG antibodies accurately, but their diagnostic performances vary with respect to the phase of the infection.


2021 ◽  
Vol 12 ◽  
Author(s):  
Mikhail Petrovich Kostinov ◽  
Pavel Ivanovich Zhuravlev ◽  
Lylia Solomonovna Gladkova ◽  
Kirill Vadimovich Mashilov ◽  
Valentina Borisovna Polishchuk ◽  
...  

It has been proven that post-vaccination immunity to measles virus after two doses of vaccine is not able to persistently protect against infection throughout life. The goal of this research was to determine the immune layer to the measles virus among women in labor and maternity ward personnel in the same medical institution. The levels of IgG antibodies to measles virus in the umbilical cord blood of 594 women in labor and 88 workers of the maternity ward were studied by ELISA. It was revealed that 22.7% of umbilical cord blood serum samples from parturient women and 21.4% of blood serum samples from maternity ward personnel were seronegative (<0.18 IU/ml). Levels of IgG antibodies to measles virus in low values (<1.0 IU/ml) were detected in 67% of blood serum samples among women in labor and 68.9% among employees of the maternity ward. Among women in labor, women under 35 years of age are at the highest risk of contracting measles; the proportion of women with low levels of protective antibodies in this age group was almost 70%, and the proportion of women without protective levels of antibodies was 23%. Compared with the age group 36–43, the age of women in labor under 35 was associated with a higher chance of not having immune protection against infection with measles virus OR [95% CI] = 2.2 [1.1–4.5] (p = 0.02) or had a low level of protection OR [95% CI] = 1.9 [1.2–3.0] (p = 0.001). It was also found that among women over 35 years of age, the proportion of persons with a high level of antibodies in women in labor was statistically significantly higher than among members of the maternity ward staff (13 and 0%, respectively, p = 0.007). Thus, maternity ward employees and women in labor constitute a risk group for measles due to the presence of a high proportion of seronegative persons among women of childbearing age (both maternity ward employees and women in labor). These conditions create the need to revise current approaches to present vaccination procedures, especially in the current epidemiological situation with COVID-19.


2020 ◽  
Vol 10 (4) ◽  
pp. 729-734
Author(s):  
V. D. Stoiljkovic ◽  
M. A. Bichurina ◽  
I. N. Lavrentieva ◽  
S. B. Filipovic-Vignjevic ◽  
M. D. Bancevic ◽  
...  

In 2017, the WHO registered 23,927 measles cases in 44 out of 53 countries in the European region. In 2018, measles incidence rate increased up to 82,599 cases registered in 48 countries of the region, with a large number of measles-associated deaths. Overall, 72 measles fatalities were registered in 10 European countries, including Serbia (15 cases).Aim of the study: to characterize 2017—2018 epidemiological upsurge of measles incidence rate observed in the Republic of Serbia (RS) and the Northwestern Federal District (NWFD) of the Russian Federation.Materials and methods. During the 2017—2018 season, 944 serum samples were collected from patients with measles, rubella, or exanthematous diseases in the NWFD and tested in the Laboratory of Virology at the St. Petersburg Regional Centre for Measles Surveillance (SPbRC). In 2017—2018, 2,946 serum samples from the Republic of Serbia were analyzed in the SPbRC by using ELISA with IgM measles test system (Vector-Best, Russia; or Siemens, Germany). Urine and swab samples were examined by RT-PCR and used for isolation and genotyping of measles viruses.Results. From 2017 to 2018, 5,798 measles cases were registered in the RS, among which 2,946 were laboratory-confirmed (serological testing and/or PCR). Unvaccinated subjects or those with unknown vaccination status accounted for majority of the cases. Children under 5 years of age and adults aged 30 years and over dominated among measles patients. During this season, 15 deaths were reported. Several genotypes of measles virus circulated in the RS, e.g. В3 Dublin, D8 Gir Somnath, and D8 Herborn. In 2018, 109 measles cases were recorded in the NWFD, 5 of which were imported from abroad. Among patients, adults comprised 64.2%, wherein 74.3% were covered by unvaccinated subjects or those with unknown vaccination status. Rise in measles incidence rate linked to multiple importations of various measles virus genotypes: В3 Kabul; B3 Dublin; D8 Frankfurt; D8 Cambridge; and D8 Gir Somnath.


2007 ◽  
Vol 5 (2) ◽  
pp. 267-282 ◽  
Author(s):  
Jeremy Olstadt ◽  
James Jay Schauer ◽  
Jon Standridge ◽  
Sharon Kluender

Since 2002, the United States Environmental Protection Agency (USEPA) has approved ten enzyme-based total coliform and E. coli detection tests for examination of drinking water. These tests include: Colilert®, Colilert-18®, Colisure®, m-Coli Blue 24®, Readycult® Coliforms 100, Chromocult®, Coliscan®, E*Colite®, Colitag™ and MI Agar. The utility of the enzyme based test systems is based on both the ability of the test to detect the target organisms at low levels and the ability of the test system to suppress the growth of non-target organisms that might result in false positive results. Differences in the ability of some of these methods to detect total coliform and E. coli, as well as suppress Aeromonas spp., a common cause of “false positive” results, have been observed. As a result, this study was undertaken to elucidate the strengths and weaknesses of each method. Water samples were collected from three geographically and chemically diverse groundwaters in Wisconsin. One-hundred milliliter aliquots were individually spiked with both low concentrations (one to ten organisms) and high concentrations (fifty to one-hundred) of each of five different total coliform organisms (Serratia, Citrobacter, Enterobacter, E. coli, & Klebsiella). These spiked samples were used to test the capability of ten enzyme-based test systems to both detect and enumerate the spiked organisms. In addition, 100 ml samples were independently spiked with two different strains of Aeromonas spp. at six different levels, to assess the ability of each enzyme-based test to suppress Aeromonas spp. Analysis of the data indicated that wide variability exists among USEPA approved tests to detect and quantify total coliforms, as well as suppress Aeromonas spp.


2021 ◽  
Vol 66 (11) ◽  
pp. 689-694
Author(s):  
A. L. Shutikova ◽  
G. N. Leonova ◽  
A. F. Popov ◽  
M. Yu. Shchelkanov

The coexistence of various pathogens inside the patient’s body is one of the poorly studied and current issues. The aim of the study is to identify the relationship between the indicators of complex laboratory diagnostics and the clinical manifestations of a mixed disease during subsequent infection with the SARS-CoV-2 virus using the example of a case of chronic encephalitis-borreliosis infection. Seven blood serum samples were collected from the patient over the course of a year. For the etiological verification of the causative agents of TBE, Lyme disease and COVID-19, the methods of ELISA and PCR diagnostics were used. The patient was diagnosed with Lyme disease on the basis of the detection of IgG antibodies to Borrelia 5 months after the onset of the disease, since she denied the tick bite. In the clinical picture, there was an articular syndrome and erythema migrans. Later, IgG antibodies to the TBEV were found in the blood. Throughout the study, IgM antibodies to Borrelia were not detected. The exacerbation of Lyme disease could be judged by the clinical manifestations of this disease and by the growth of specific IgG antibodies. A feature of this case was that during an exacerbation of the Lyme disease, an infection with the SARS-CoV-2 virus occurred. Treatment (umifenovir, hydroxychloroquine, azithromycin, ceftriaxone) was prescribed, which improved the condition of the underlying disease, decreased joint pain, decreased IgG levels to borrelia. However, during this period, serological markers of TBEV appear: antigen, IgM antibodies, and the titer of IgG antibodies increases. Most likely, this was facilitated by the switching of the immune system to the SARS-CoV-2 virus, with the simultaneous suppression of borrelia with antibiotics and the appointment of hydroxychloroquine, which has an immunosuppressive effect. Despite the activation of the virus, clinical manifestations of TBE were not observed in the patient, which is most likely associated with infection with a weakly virulent TBEV strain. The further course of tick-borne infections revealed the dominant influence of B. burgdorferi in relation to TBEV. Laboratory studies have shown that suppression of the activity of the borreliosis process by etiotropic treatment subsequently led to the activation of the persistent TBEV.


2019 ◽  
Vol 64 (9) ◽  
pp. 553-559
Author(s):  
N. V. Rudakov ◽  
S. V. Shtrek ◽  
A. I. Blokh ◽  
N. A. Penjevskaya ◽  
L. D. Shchuchinova

The real epidemiological impact of Spotted Fever Group rickettsioses including Siberian tick-borne typhus (STT) in Russia is not sufficiently studied. One of the reasons is the actual absence of either certified domestic diagnostic kits or the evidence for using foreign test kits for laboratory verification of this group of tick-borne infections in medical practice. Objective of our study was to study the diagnostic accuracy of the ELISA test system based on Rickettsia conorii antigens for serological verification of STT. The ROC analysis was performed and operational characteristics (sensitivity, specificity, accuracy, likelihood ratio of positive and negative results) of the STT serological verification test to identify IgM to rickettsia at different times from the onset of the disease using a test system to detect antibodies to Rickettsia conorii were calculated based on the results of a survey of two groups of patients comparable by gender and age (34 patients with pathognomonic signs of STT and 76 clinically healthy people). It was found that the detection of IgM antibodies to rickettsia using the Rickettsia conorii IgM/IgG ELISA test system (Vircell) allows the disease to be verified 10-14 days after the onset of clinical symptoms in 72% (56-88%) of STT patients. We recommend the interpretation of results of the test system “Rickettsia conorii ELISA IgM/IgG” for serological verification of STT which differ from the manufacturer’s recommendations regarding verification of Mediterranean fever caused by R. conorii in the following way: the diagnosis of STT should be considered laboratory confirmed when the index of IgM antibodies (IAT) exceeds 8.0; if the IAT is less than 5.0 then a repeated examination of the patient after 10-14 days will be necessary; if the IAT is in the range of 5.0-8.0 then the sample should be re-examined and / or the patient should be examined after 10-14 days. The use of the test system “Rickettsia conorii ELISA IgM / IgG” is promising for laboratory diagnosis and seroepidemiological studies of Spotted Fever Group rickettsioses in Russia.


Folia Medica ◽  
2016 ◽  
Vol 58 (4) ◽  
pp. 250-256 ◽  
Author(s):  
Stefka Kr. Ivanova ◽  
Svetla G. Angelova ◽  
Asya P. Stoyanova ◽  
Irina L. Georgieva ◽  
Lubomira K. Nikolaeva-Glomb ◽  
...  

AbstractBackground:Inflammatory diseases of the heart (myocarditis, pericarditis) are commonly caused by viruses. Among the human cardiotropic viruses, parvovirus B19, Coxsackie B viruses, and adenoviruses play a leading role.Aim:The aim of the present study was to determine the presumptive causative role of parvovirus B19, Coxsackie B viruses, and adenoviruses in the development of myocarditis, pericarditis and dilated cardiomyopathy by demonstrating the presence of specific antiviral antibodies or viral DNA in patients’ serum samples.Materials and methods:We tested serum samples collected between 2010 and 2014 from 235 patients with myocarditis (n=108), pericarditis (n=79), myopericarditis (n=19), dilated cardiomyopathy (n=7), and fever of unknown origin accompanied by cardiac complaints (n=22). The mean age of patients with the standard deviation was 33 ± 18 years. Serological and molecular methods (ELISA for specific IgM/IgG antibodies to parvovirus B19 and IgM antibodies to Coxsackie B viruses and adenoviruses, and PCR for detection of parvovirus B19 in serum samples, respectively) were used in the study.Results:Of all tested 235 serum samples, in 60 (25.5%) positive results for at least one of the three tested viruses were detected. Forty out of these 235 serum samples (17%) were Coxsackie B virus IgM positive. They were found in 17% (18/108) of the patients with myocarditis, in 15% (12/79) of those with pericarditis, in 16% (3/19) of those with myopericarditis and in 32% (7/22) in those with fever of unknown origin. The 63 Coxsackie B virus IgM negative patient’s serum samples were tested by ELISA for presence of adenovirus IgM antibodies. Such were found in 4 patients with pericarditis and in 2 patients with fever of unknown origin. Every IgM negative sample (n=189) for Coxsackie B and adenovirus was further tested by ELISA for parvovirus B19 IgM/IgG antibodies. B19-IgM antibodies were detected in 14 patients (7.4%). The percentages for B19-IgM antibodies was 8% (7/90), 5% (3/63) and 31% (4/13) in the patients affected with myocarditis, pericarditis, and fever of unknown origin, respectively. Protective B19-IgG antibodies were found in 108 (57%) of the samples. A B19-PCR signal was detected in all the patients who were B19-IgM positive, and in only 1 patient with positive B19-IgG result, the latter presenting with dilated cardiomyopathy.Conclusion:The present study shows the involvement of Coxsackie B, parvovirus B19 and adenoviruses in the development of inflammatory diseases of the heart (myocarditis and pericarditis). It is the first ever study in the country that simultaneously analyzes the prevalence of the three major human cardiotropic viruses.


Author(s):  
Flywell Kawonga ◽  
Gerald Misinzo ◽  
Dylo Pemba ◽  
Leonard Mboera ◽  
Isaac Thom Shawa

Chikungunya is a mosquito-borne viral disease caused by Chikungunya virus (CHIKV. We conducted this study determine the seroprevalence and clinical presentation of Chikungunya infection among outpatients seeking healthcare in Mzuzu City, Malawi. Blood samples were collected from malaria negative and non-septic febrile outpatients with fevers ≥38 °C, for not more than 5 days. The enzyme- linked immunosorbent assay (ELISA) test was used to detect anti-CHIKV IgM antibodies and its results were used to determine seroprevalence of Chikungunya. A total of 119 serum samples were tested, of these, 73 (61.3%) tested positive for anti-CHIKV IgM antibodies by ELISA. Laboratory requisition forms were used to capture demographic information such as age, sex, clinical signs and symptoms presented by the enrolled patients. Age groups of 1-9, 10- 19, 20- 29, 30- 39, 40- 49, and ≥50 years had 17.8% (n= 13), 12.3 %,( n=9), 15.1%) (n=11), 19.2%; (n=14), 17.8% (n=13) and 17.8% (n=13) proportion of seroprevalence respectively. Most of the CHIKV infected individuals presented with fever (52.05%), joint pain (45.21%) and abdominal pain (42.67%). The presence of anti- CHIKV IgM antibodies suggest the presence of recent CHIKV infection and therefore accurate laboratory assays are highly recommended for CHIKV diagnosis and appropriate management of febrile patients.


2019 ◽  
Vol 36 (5) ◽  
pp. 35-43
Author(s):  
Dmitriy Yu. Sosnin ◽  
Ol'ga Yu. Nenasheva ◽  
Nadezhda A. Zubareva ◽  
Andrey V. Renzhin ◽  
Konstantin R. Gal'kovich

Aim. Blood serum and urinary procalcitonin (PCT) concentration in healthy persons was studied. Materials and methods. A single-stage observational study of case-control type was performed. The study included 32 men and 37 women of middle age (53.4 16.4 years) with normal renal function. PTC concentration was determined with the method of solid-phase enzyme immunoassay using test-system (Procalcitonin IFA-BEST, Russia). Results. Blood serum PCT concentration in the examined persons was 0.029 0.016 ng/ml (M SD). The number of blood serum samples with PTC level 0.05 ng/ml was 5.8 % (4 from 69). The mean urinary PTC concentration by 72.59 times exceeded the mean blood serum PTC content and was 2.12 1.832 ng/ml (р 0.000001). Coefficient of variation of results for the urine by 1.57 times exceeded the analogous index for the blood serum. When comparing the results of blood serum and urine analyses, no statistically significant differences between men and women were revealed. When estimating the coefficients of linear correlation between PTC content in the blood serum and urine, a weak positive dependence was established (R = 0.302782). Conclusions. High PTC concentration in the urine permits to suppose that one of the ways of procalcitonin removal from the blood plasma is its elimination by kidneys in unchanged type by means of glomerular filtration.


PLoS ONE ◽  
2021 ◽  
Vol 16 (1) ◽  
pp. e0245701
Author(s):  
Sergio Estrada-Martinez ◽  
Alma Rosa Pérez-Álamos ◽  
Melina Ibarra-Segovia ◽  
Isabel Beristaín-Garcia ◽  
Agar Ramos-Nevárez ◽  
...  

The seroepidemiology of infection with Toxoplasma gondii (T. gondii) in alcohol consumers is largely undeveloped. In light of this, we sought to determine the seroprevalence of T. gondii infection in alcohol consumers in Durango, Mexico, and the association of T. gondii seroprevalence with characteristics of the population studied. Anti-T. gondii IgG and IgM antibodies were searched in sera of participants using commercially available enzyme immunoassays. Bivariate and logistic regression analyses were then used to determine the association between T. gondii infection and the characteristics of the population studied. Of the 1544 people studied (mean age: 39.4±14.0 years), 173 (11.2%) tested positive for anti-T. gondii IgG antibodies. We were able to test 167 of the 173 anti-T. gondii IgG positive sera for anti-T. gondii IgM antibodies. Fifty-five (32.9%) of these 167 serum samples were positive for anti-T. gondii IgM antibodies. Bivariate analysis showed that visual impairment, history of surgery, and hepatitis were negatively associated with T. gondii IgG seropositivity (P<0.05). In women, seropositivity to T. gondii was positively associated with a history of pregnancy (P<0.05). Logistic regression analysis showed that T. gondii seropositivity was associated with the variables consumption of armadillo meat (OR = 2.33; 95% CI: 1.04–5.22; P = 0.03), and the use of latrines for elimination of excretes (OR = 2.27; 95% CI: 1.07–4.80; P = 0.03); and high (>150 IU/ml) anti-T. gondii IgG antibodies were associated with consumption of both armadillo meat (OR = 2.25; 95% CI: 1.01–5.02; P = 0.04) and crowding at home (OR = 1.63; 95% CI: 1.02–2.61; P = 0.03). We found a distinct T. gondii seroprevalence in people with alcohol consumption from those previously found in population groups in the region. This is the first study that illustrates the association between high anti-T. gondii antibodies and crowding in Mexico, and the second study on the association between T. gondii infection and consumption of armadillo meat and the use of latrines in this country. We conclude that epidemiology of T. gondii infection in people with alcohol consumption deserves further investigation.


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