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Tetanus antitoxin is quantified in Cuba at blood banks, from the serum of immunized donors, to produce aspecific human gamma globulin. A heterogeneous indirect immunoenzymatic assay is used, using the serum as analytical sample. The possible use of plasma obtained from plasmapheresis as alternative sample was evaluated in this research, to minimize the volume of total blood extracted to the donors. One hundred plasma donors who came to donate between October and November 2013 were selected by simple random sampling. Serum sample was obtained for extraction of 5 mL of blood, deposited in dry glass tube. While the other sample took 1.5 mL of plasma in a plastic tube with cover, at the end of the donation directly of the unit of plasma collected. Comparison of the difference between the means of both groups was done using SPSS for Windows. It was found that the values obtained in serum were bigger than those obtained in plasma. Difference between the means of both groups was statistically significant (p 0.00). It is not advisable to use the obtained plasma of the plasmapheresis as analytic sample in this assay.

Para el diagnóstico del virus de la hepatitis C, se utilizan ampliamente las pruebas de Elisa por su sensibilidad y especificidad. Gran parte de ellos se basa en el empleo de péptidos sintéticos. Una de las zonas más conservadas y de gran antigenicidad es la región estructural del virus. En este estudio, se sintetizaron siete péptidos de zonas informadas como altamente antigénicas de la región estructural del virus de la hepatitis C. Los péptidos sintetizados representan la región del núcleo del virus. Se realizó la evaluación y comparación de los resultades de los péptidos sintetizados, para lo cual se emplearon muestras positivas (n=72) y negativas (n=42). Con los péptidos más cercanos a la región amino terminal, la reactividad fue alta, pero fue disminuyendo a medida que nos acercábamos a la región carboxilo terminal. Estos resultados coinciden con lo informado por otros autores.

Para el diagnóstico del virus de la hepatitis C, se utilizan las pruebas de Elisa, debido a su sensibilidad y especificidad. Gran parte de estas pruebas se basa en el empleo de péptidos sintéticos y, en la actualidad, de péptidos quiméricos. En este estudio se sintetizó un péptido quimérico que comprende secuencias inmunodominantes de las regiones no estructurales NS4 y NS5 del virus de la hepatitis C. El péptido representativo de las dos secuencias está separado por un brazo espaciador de dos residuos de glicina. El péptido se evaluó como antígeno en un ensayo UMELISA, utilizando muestras positivas (n=30), muestras negativas (n=40) y el panel de Boston Biomedica Inc. (n=40). Los resultados del péptido quimérico NS41 NS5 se compararon con los resultados de los péptidos individuales NS4 y NS5 y con la mezcla de estos dos péptidos. Los resultados de los péptidos individuales coinciden con los datos informados en la literatura. Con la mezcla de los péptidos, no aumentó la detección de muestras positivas, pero con el péptido quimérico se logró aumentar la sensibilidad.

Tetanus antitoxin is quantified in Cuba at blood banks, from the serum of immunized donors, to produce a specific human gamma globulin. A heterogeneous indirect immunoenzymatic assay is used, using the serum as analytical sample. The possible use of plasma obtained from plasmapheresis as alternative sample was evaluated in this research, to minimize the volume of total blood extracted to the donors. One hundred plasma donors who came to donate between October and November 2013 were selected by simple random sampling. Serum sample was obtained for extraction of 5 mL of blood, deposited in dry glass tube. While the other sample took 1.5 mL of plasma in a plastic tube with cover, at the end of the donation directly of the unit of plasma collected. Comparison of the difference between the means of both groups was done using SPSS for Windows. It was found that the values obtained in serum were bigger than those obtained in plasma. Difference between the means of both groups was statistically significant (p 0.00). It is not advisable to use the obtained plasma of the plasmapheresis as analytic sample in this assay. [ABSTRACT FROM AUTHOR]

The use of dry blood spots (DBS) on filter paper has become of increasing interest in the field of immunoassay over the last years due to the advantages this mode of presentation brings, enabling its use in countries where resources are scarce or limited to accomplish epidemiological studies. In this work, the optimal conditions for the elution of dry blood spots (3 mm disks) collected on filter paper were determined. Serum samples and the elution conditions establish for 5 mm disks were taken as reference in the detection of specific antibodies to Trypanosoma cruzi by using the indirect UMELISA® CHAGAS (International Tecnosuma S.A.) immunoassay based on Ultra Micro Analytic System (SUMA®). Twelve sero-positive serum samples for Chagas disease from Colombia were confirmed by indirect immunofluorescence (IFI) and ELISA -- Chagatek®, and 75 DBS on filter paper from Bolivia reactive by UMELISA® CHAGAS were evaluated. Additionally, serum samples and DBS negative to Chagas disease from healthy donors from Havana Province Blood Bank were tested. The sensitivity and specificity study results for 3 mm and 5 mm disks were similar. Reduction of the disk diameter allows increasing the number of diagnostic tests, from a very same spot, for the detection of other disease such as the acquired immunodeficiency syndrome, hepatitis B and C, among others. [ABSTRACT FROM AUTHOR]

• PSA detection and quantification is used as a tumor marker. • UMELISA PSA is an assay for measurement of total and free PSA in human serum samples. • Two-in-one test kit capable of quantifying and differentiating total and free PSA. • Fluorescent assay that uses only 10 µL of sample and reagents. • Cost-effective diagnostic kit implemented in the Cuban Public Health System. [Display omitted] Prostate specific antigen (PSA) is a serine protease used for the screening of prostate cancer. The total portion of PSA (tPSA) can be found in its free form (fPSA), or bound to other proteins forming a stable complex. A heterogeneous sandwich-type UltraMicro Enzyme-Linked ImmunoSorbent Assay (UMELISA) has been developed for the measurement of tPSA and fPSA in human serum samples. Strips coated with a high affinity monoclonal antibody (MAb) directed against PSA are used as solid phase, to ensure the specificity of the assay. Biotinylated MAbs specific for tPSA and fPSA ensured sensitivity, given the high affinity binding to streptavidin. The assay was completed in 1.5 h, with a measuring range 0.019–20 µg/L (tPSA), and 0.009–20 µg/L (fPSA). The intra- and inter-assay CV were lower than 9%. Recovery percentages were 96–105%. High correlations were found between the values of the UMELISA PSA standards and the International Reference Standards 96/670 (R2 = 0.9996) and 96/688 (R2 = 0.9989). The assay did not recognize any of the interfering molecules tested. Regression analysis of serum samples showed a good correlation with Roche Elecsys total PSA (n = 631, R2 = 0.986, ρc = 0.992), BioMérieux VIDAS TPSA (n = 631, R2 = 0.989, ρc = 0.993) and Roche Elecsys free PSA (n = 164, R2 = 0.973, ρc = 0.979), all with a relative difference below 15%, and a p < 0.001. A retrospective study of the use of UMELISA PSA in Cuba was carried out. The analytical performance characteristics of UMELISA PSA support its use for the quantification of tPSA and fPSA in human serum samples in a single kit, making it an affordable diagnostic assay available to Cuban Public Health System and developing countries. Between the years 2014–2020, more than 3 million Cuban patients have benefited from the test for free. [ABSTRACT FROM AUTHOR]

Abstract: Six chimeric synthetic peptides (QCha-1, QCha-2, QCha-3, QCha-4, QCha-5, and QCha-6) incorporating antigenic sequences of two immunodominant repeat B-cell epitopes of Trypanosoma cruzi were synthesized by conventional solid-phase peptide synthesis. The antigenic activity of these peptides was evaluated by UltramicroEnzyme-linked immunosorbent assay (UMELISA) by using panels of positive Chagasic sera (n =82), while specificity was evaluated with samples from healthy blood donors (n =44) and patients with other infectious diseases (n =86). The antigenicity of the chimeric peptides in solid-phase immunoassays was compared with that of the monomeric peptides. Data demonstrated that the chimeric peptide QCha-5 was the most reactive because it detected antibodies to parasite efficiently. The results indicate that chimeric peptide as coating antigen is very useful for the immunodiagnosis of Chagas’ disease. [Copyright &y& Elsevier]

Abstract: Four chimeric synthetic peptides (Q5, Q6, Q7⊗, and Q8⊗), incorporating immunodominant epitopes of the core p19 (105–124a.a.) and envelope gp46 proteins (175–205a.a.), of HTLV-I were obtained. Also, two gp46 monomeric peptides M4 and M5⊗ (Ser at position 192) were synthesized. The analysis of the influence of the peptide lengths and the proline to serine substitution on the chimeric and monomeric peptides’ antigenicity, with regard to the chimeric peptides Q1, Q2, Q3⊗, and Q4⊗, reported previously, for HTLV-I was carried out. The peptides’ antigenicity was evaluated in an ultramicroenzyme-linked immunosorbent assay (UMELISA) using sera of HTLV-I/II. The peptides’ antigenicity was affected appreciably by the change of the peptide length and amino acid substitutions into the immunodominant sequence of gp46 peptide. [Copyright &y& Elsevier]

The antigenicity of three chimeric synthetic peptides (Qm, Qm-16, and Qm-17) incorporating an immunodominant epitope of the gp41 transmembrane protein (587–617) and the different epitopes of the gp120 envelope protein (495–516), (301–335), (502–516) of human immunodeficiency virus (HIV-1), separated by two glycine residues, was evaluated by UltramicroEnzyme-linked immunosorbent assay (UMELISA) by using panels of anti-HIV-1 positive sera (n = 47). The specificity was evaluated with samples from healthy blood donors (n = 20) and anti-HIV-2 positive samples (n = 10). The results indicate that the chimeric peptide, Qm, was the most reactive one because it detected antibodies to virus efficiently. This may be related to peptide adsorption onto the solid surface, the C-terminal region of HIV-1 gp120 (495–516) combined with gp41 (587–617) in the chimera, and the epitope accessibility to the antibodies. This study showed the usefulness of the chimeric peptides as antigen to detect antibodies to HIV-1 virus. [ABSTRACT FROM AUTHOR]

Monomeric and chimeric synthetic peptides were used as coating antigens in four different mixtures in a solid phase immunoassay to select an optimal combination for the detection of antibodies to human T-cell lymphotropic virus (HTLV) in serum samples. The peptides, P-13 (gp21 I), Q5 (gp21 II)-GG-(gp46 II), and Q (gp46 I)-GG-(p19 I), represent immunodominant sequences from transmembrane protein (gp21), envelope protein (gp46), and core protein (p19) of HTLV I/II viruses; they were the most antigenic and specific peptides in previous studies. The sequences of the chimeric peptides were separated by two glycine residues. An indirect UltramicroEnzyme-linked immunosorbent assay (UMELISA) was used to evaluate the antigenicity of these peptide mixtures by using samples from anti-HTLV I/II PRP205(M), (n = 20), HTLV I-infected individuals from Cuba (n = 7), and HTLV I-positive sera from Colombia and Chile (n = 9). The specificity was evaluated with healthy blood donor sera (n = 300), anti-HIV 1-positive samples (n = 10), and other seropositive samples to different infectious agents. The highest sensitivity and specificity was obtained with mixture 1, which could be very useful in the immunodiagnostic of HTLV infection. [ABSTRACT FROM AUTHOR]

A chimeric synthetic peptide incorporating immunodominant epitope of the p19 gag protein (116–134) and the gp46 env protein (178–200) of HTLV-II virus, separated by two glycine residues, was synthesized by conventional solid-phase peptide synthesis. The antigenic activity of this peptide was evaluated by Ultramicro Enzyme-linked immunosorbent assay (UMELISA) by using panels of anti-HTLV-II positive sera (n = 9), anti-HTLV-I/II positive sera (n = 2), HTLV-positive (untypeable) serum samples (n = 1), and anti-HTLV-I positive sera (n = 14), while specificity was evaluated with samples from healthy blood donors (n = 20). The efficacy of the chimeric peptide in solid-phase immunoassays was compared with the monomeric peptides. Data demonstrated that the chimeric peptide was the most reactive because it detected antibodies to virus efficiently. This may be related to peptide adsorption to the solid surface and epitope accessibility to the antibodies. The results indicate that chimeric peptide as coating antigen is very useful for the immunodiagnosis of HTLV-II infection. [ABSTRACT FROM AUTHOR]

Congenital hypothyroidism (CH) is one of the most common preventable causes of mental retardation. Newborn screening and thyroid therapy started within 2 weeks of age can normalize the cognitive and motor impairment caused by lack of thyroid hormone during the early postnatal phase of brain development. The availability of assays to determine thyroxin (T4) and thyroid stimulating hormone has allowed the establishment of newborn screening programs for CH. The Immunoassay Center (Cuba) developed the T4 Neonatal UMELISA(r) to determine neonatal T4 levels in dried blood on filter paper using 5 mm discs of samples, standards and controls. The number of diseases which can be diagnosed in Neonatal Screening Programs has increased, so it is necessary to make the maximum use of blood samples collected on filter paper. This work shows the standardization of the T4 Neonatal UMELISA(r) modified in order to use 3 mm discs. The assay was completed in 3 1/2 hours, with a measuring range of 25-400 nmol/L. The intra- and inter-assay coefficients of variation (CV) were 6-10 % and 7-12 % respectively, depending on the T4 concentrations. The recovery ranged from 91.8-115.1 %. The modified assay showed high Pearson correlation (r = 0.877) and concordance correlation (?c = 0.867) with the T4 Neonatal UMELISA(r). The performed modifications do not affect the sensitivity, precision and accuracy of the assay, permitting a more rational use of newborn blood samples and the possibility to increase the number of diseases included in the newborn screening programs.