Evaluation of rapid, cassette immunochromatographic tests in the serological diagnosis of COVID-19

Introduction: Lateral flow assays (LFIA) are the technology behind low-cost, simple, rapid and portable detection devices popular in biomedicine. Lately, they are very common used in serodiagnosis of SARS-CoV-2 infections. The aim of the presented study was to assess the usefulness of selected LFIA in serological diagnosis of COVID-19. Methods: The usefulness of seven lateral flow assays in the serodiagnosis of COVID-19 was evaluated (VAZYME, DIAGNOSIS, PCL, INGEZIM, BIOSENSOR, ACCU-TELL, NOVAtest). The study used 107 serum samples obtained from 74 individuals with current SARS-CoV-2 infection confirmed by RT-PCR. The ELISA-IgG (Euroimmun) was used as the reference assay for sensitivity and specificity testing. Results: The highest percentage of positive results was obtained when searching for IgG antibodies with the NOVAtest (40.6%) and DIAGNOSIS (39.2%) sets and the lowest detection for the PCL set - 25.5%. In the case of searching for IgM antibodies in all sets, significantly lower percentages of positive results compared to the IgG class were recorded. In general, all lateral flow assays showed low sensitivity in relation to the Euroimmun ELISA-IgG. The DIAGNOSIS kit (64.5%) was characterized by the highest sensitivity, and the PCL kit was the lowest (38.7%). On the other hand, the specificity of all kits was very high, almost 100% in almost all cases. Conclusions: Lateral flow assays due to their low sensitivity are not suitable for quick diagnosis of the current SARS-CoV-2 infections and cannot be an alternative to genetic or even antigen tests. They may be used only to retrospectively test the presence of IgG antibodies. However, a negative results of LFIA in suspected disease or after vaccination should be confirmed by more sensitive serological tests.

Comparative Study of Indirect Fluorescent Antibody, ELISA, and Immunochromatography Tests for Serological Diagnosis of Bovine Babesiosis Caused by Babesia bovis

The indirect fluorescent antibody test (IFAT) is the most frequently used test to conduct seroepidemiological studies so far, and it is regarded as the "gold standard" test for the serological diagnosis of bovine babesiosis. The aim of the present study was to compare the enzyme-linked immunosorbent assay (ELISA) and the rapid immunochromatography test (ICT) for use in the serological diagnosis of cattle exposed to B. bovis in Mexico. The evaluation of test performance was carried out with 30 positive and 30 negative reference sera. A total of 72 bovine sera samples collected from cattle in a region with endemic bovine babesiosis were analyzed by ELISA and ICT, and the results were compared with those of IFAT. Kappa value (k) was also calculated to determine the agreement between tests. The sensitivity and specificity of ELISA for detecting antibodies against B. bovis were 87% (26/30) and 80% (24/30), respectively. The sensitivity and specificity of ICT for detecting antibodies against B. bovis were 90% (27/30) and 83.3% (25/30), respectively. The overall concordance determined for ELISA and ICT was 94.4% (68/72) and 98.6% (71/72), respectively, when the results were compared with those of IFAT. ICT was more sensitive and specific in this comparative study, showing good strength of agreement (k = 0.79) with respect to IFAT. ICT combines a strip-based assay system that is fast, practical, and sensitive for detection of antibodies to B. bovis, which suggests that it could be applied in the field without requiring any laboratory equipment for its use and interpretation of test results.

Immunoreactivity of Polish Lyme Disease Patient Sera to Specific Borrelia Antigens—Part 1

The diverse clinical picture and the non-specificity of symptoms in Lyme disease (LD) require the implementation of effective diagnostics, which should take into account the heterogeneity of Borrelia antigens. According to available guidelines, laboratories should use a two-tier serological diagnosis based on the enzyme-linked immunosorbent (ELISA) screening test and confirmation of the immunoblot (IB). The aim of the study was to investigate the immunoreactivity of LD patient sera to Borrelia antigens and to attempt to identify the genospecies responsible for LD using an ELISA–IB assay combination. Eighty patients with suspected LD and 22 healthy people participated in the study. All samples were tested with ELISA and IB assays in both IgM and IgG antibodies. In the case of the ELISA assay, more positive results were obtained in the IgM class than in the IgG class. In the case of the IB assay, positive results dominated in the IgG class. Positive results obtained in the IB assay most often showed IgM antibodies against the OspC and flagellin antigens, whereas the IgG antibodies were against VlsE, BmpA, OspC, p41, and p83 antigens. The IB assay is an important part of LD serodiagnosis and should be mandatory in diagnostic laboratories.

Are humanized IgE reporter systems potential game changers in serological diagnosis of human parasitic infection?

Immunoglobulin E (IgE) is thought to have evolved to protect mammalian hosts against parasitic infections or toxins and plays a central role in the pathogenesis, diagnosis, and therapy of IgE-mediated allergy. Despite the prominence of IgE responses in most parasitic infections, and in stark contrast to its use in the diagnosis of allergy, this isotype is almost completely unexploited for parasite diagnosis. Here, we discuss the perceived or real limitations of IgE-based diagnosis in parasitology and suggest that the recent creation of a new generation of very sensitive cellular IgE-based reporters may represent a powerful new diagnostic platform, but needs to be based on a very careful choice of diagnostic allergens.

Serological diagnosis of gonorrhea

Alf. Cohn (Derm. Ztschr. Bd. 55, H. 2) gives detailed instructions for the preparation of a specific antigen, patient serum, and also describes in detail the technique of the Bordet-Gengou reaction itself. The author received as a result of his observations with acute gonorrhea in men 40% pos. responses, for chronic 60% and for complicated 80-100%. The results are especially demonstrative in gonorrhoid arthritis (100%). It is important to use the reaction in doubtful cases, when microscopic and bacteriological examinations do not give the necessary answer for making a diagnosis. If people who do not have gonorrhea are vaccinated, the reaction may be positive. If a positive reaction persists for months or years, you need to look for a focus with gonococci. According to the author, the complement fixation reaction cannot replace the previously tested diagnostic methods, but it complements them. Together with bacteriological and clinical research methods, the reaction sometimes helps to understand the diagnosis of difficult cases.

Clinical observations of serological diagnosis of gonorrhea

Carl Funk (Derm. Ztschr. Bd. 55, H. 2, 1929) performed a complement rejection reaction in 600 gonorroics and in 100 people with various other diseases, and in acute gonorrhea A. received 60% of positive results, in chronic - 80%, with epididymitis 90%, with prostatitis and spermatocystitis 97%, with arthritis, bursitis and tendovaginitis 97%. The brightness of the reaction is observed on the 14th day after the onset of the disease. According to the author's observations, the reaction is characterized by great specificity, only in isolated cases (polyart. Reumatica) there is a nonspecific delay in hemolysis. The reaction is of great service in differentiating doubtful cases of inflammation of the appendages, eyes, joints, heart disease and gonococcal sepsis and a number of other diseases of the genitourinary sphere. Stable positive, the reaction of deviation of complement in cases of establishing the fact that gonorrhea has been cured indicates the presence of a hidden focus with gonococci.

Comparison of different detection methods for Ascaris suum infection on Austrian swine farms

Background Ascaris suum, the large roundworm of pigs, is one of the economically most important pig parasites worldwide. In Austria it is commonly diagnosed by monitoring livers for milk spots at the slaughterhouse and intravital diagnosis (flotation for detection of fecal egg shedding). Recently, serological diagnosis based on the detection of specific antibodies with an ELISA (SERASCA®) with high sensitivity has been developed. To introduce and evaluate serology for A. suum screening in Austrian pigs, blood (for serology) (n = 177) and feces (for copromicroscopy) (n = 177) were taken from randomly selected slaughter pig batches from 18 farms at a slaughterhouse in Lower Austria. In addition, livers presented at slaughter (n = 844; max. 70/farm) were evaluated for milk spots. Results Overall, 19% of the livers were milk spot-positive (22% of those with complete diagnostic evaluations). Thirteen percent of the fecal samples contained A. suum eggs, while 69% of the blood samples were serologically positive. Despite we did not determine the sensitivity of the ELISA specifically, results ouf our study confirmed the high sensitivity of the ELISA, which was claimed by the manufacturer prior to our work (sensitivity: liver assessment: 23.5–27.0%; copromicroscopy: 8.5–9.0%; ELISA: 99.5%), and a high percentage of A. suum infections that remained undetected by standard liver assessment. Conclusions This suggests that the current method of roundworm diagnostics is insufficient and antibody detection at the end of the fattening period should be established as the standard procedure.

A comparative study of serological diagnosis of Dengue outbreak 2019

Purpose: The interpretation & correlation of the different laboratory parameters in positive dengue cases in order to eval- uate that which laboratory test is more significant for diagnosis of Dengue. Methods: Prospective examination of samples (patients’ serum) for dengue virus of different genotype by using multiplex anti-dengue IgM, IgG. We have done NS-1 test by (ICT) immunochromatographic devices, and complete blood picture (CBC) by Sysmex XP-100. Result: Detection of Viral RNA in 100 patients showed effects in the total of 73 (73.0%) samples. This graphical compar- ison shows the whole positive cases including dengue NS-I antigen, dengue serology (IgM & IgG), total 62 positive cases of NS-I are detected, 10 positive cases of dengue IgM and 9 positive cases of IgG detected, in which Complete Blood Test (CBC) shows remarkable reduction in Platelets (32 cases) and Leucopenia in (24 positive cases). Conclusion: In this research, it is concluded that the diagnosis of dengue cases is preliminary limited to initial stages i.e. CBC or sometimes dengue NS-I, as dengue IgM severity is more effective than that of Dengue NS-I & IgG. Many patients who had negative results in CBC and NS-1 testing, became positive when IgM and IgG serology testing has been done. Keywords: Dengue; NS-1; IgG; IgM; Immunochrometographic test.