The Use of Immunochromatographic Technique for Rotavirus Detection: Experience from a Tertiary Care Hospital in Central Greece

Despite the significant medical advances which have taken place in the last decades, acute diarrhoea cases remain a public health issue of major significance, with gastroenteritis agents being associated with severe symptoms in adults and high morbidity in infants and children. Regarding rotaviruses, while children are the predominant victims of rotavirus infection, adults (often caretakers or parents of these children) may experience the same symptoms of fever, vomiting, and non-bloody diarrhoea. Three different routine schemes for the detection of rotaviruses in archived stool samples were evaluated in terms of diagnostic performance. A total of 640 archived stool samples were included in the study. The samples were screened with three different techniques: A commercial rapid immunochromatographic test, a modified in-house conventional one-step reverse-transcription polymerase chain reaction (PCR) screen protocol, and a com-mer-cial one-step real-time PCR kit. Technical aspects and considerations are discussed. © 2019 S. Karger AG, Basel. All rights reserved

Validation of fluorescence polarization assay (FPA) and comparison with other tests used for diagnosis of B. melitensis infection in sheep

Fluorescence polarization assay (FPA) is a new test for the serological diagnosis of Brucella spp. infection in animals. The FPA is validated for the diagnosis of B. melitensis infection in sheep. For this purpose, 166 sera originated from natural infected sheep (verified by culture) and 851 sera originated from healthy animals (reared in areas where B. melitensis was never been isolated) were tested. The optimum cut-off value that offers the highest diagnostic sensitivity (DSn) and diagnostic specificity (DSp) was determined at 87 mP with the use of ROC analysis. The DSn and DSp of FPA using this cut-off value are calculated at 97.6 and 98.9% with a 95% confidence interval (CI) of 93.9-99.3% and 98.0-99.5%, respectively. The DSn and DSp of FPA have been assessed also using as positive reference (n = 587), sera that gave positive results at least in two tests used for diagnosis of B. melitensis in sheep as Rose Bengal Test (RBT), modified Rose Bengal Test (m-RBT), complement fixation test (CFT), indirect Elisa (i-Elisa) and competition Elisa (c-Elisa) originated from animals reared in flocks infected by B. melitensis. The optimum cut-off value using the above panel of positive reference sera was the same offering a DSn of 95.9% with a 95% Cl, 94.0-97.4%, since the DSp remains the same. The DSn and DSp as well as performance, accuracy and agreement of FPA's result were compared with those of other tests used. The accuracy of FPA is very high, similar with that of i-Elisa. FPA is a promising assay, which offers a DSn and accuracy better that of those of the tests currently approved for the diagnosis of B. melitensis in sheep and goats. Due to its simplicity, the sort time that results can be obtained and its accuracy it can be used and improve the laboratory testing capacity as well as the efficacy of the eradication program based on test-and-slaughter policy. (c) 2005 Elsevier B.V. All rights reserved

Validation of a fluorescence polarization assay (FPA) performed in microplates and comparison with other tests used for diagnosing B-melitensis infection in sheep and goats

Fluorescence polarization assay (FPA) is a relatively new test for the serological diagnosis of Brucella spp. infection in animals. FPA, carried out in 96-well microplate format, was validated here for diagnosing B. melitensis infection in sheep and goats. This study included sera from 1933 sheep and goats, from animals reared in naturally infected flocks (verified by culture) and showing a positive reaction to two different tests conducted in parallel. In addition, 2154 sera originating from healthy sheep and goats, reared in areas where B. melitensis had never been isolated, were assayed. The optimum cut-off value offering the highest diagnostic sensitivity (DSn) and diagnostic specificity (DSp) was determined at 15 mP over the mean value of the buffer control used in each microplate as determined by receiver operating characteristic analysis. The DSn and DSp of the FPA for small ruminants carried out in microplates at this cut-off value were calculated to be 95.9% and 97.9% with 95% confidence intervals (95% CI) of 94.9-97.7% and 97.2-98.4%, respectively. The accuracy of the FPA, as expressed by determination of the area under the curve, was 0.991. Indirect ELISA and FPA tests offered the highest DSn when compared with the Rose Bengal test, the complement fixation test, the modified Rose Bengal test and competitive ELISA. The parallel or serial combination of FPA with indirect ELISA offers the highest DSn and DSp. As temperature can affect the results of the FPA, all reagents must be at the same temperature and the standard for comparison must always be read under the same conditions as the sera under test. FPA performed in microplates is a promising assay; the DSn and accuracy are better than those of the tests currently approved for diagnosing B. melitensis in small ruminants. Because of its simplicity, speed, and accuracy, this test can improve capacity for laboratory testing and the efficacy of an eradication program based on a test-and-slaughter policy. (c) 2006 Elsevier B.V. All rights reserved

Groundwater quality and location of productive activities in the region of Thessaly (Greece)

In the present study the involvement of human activities is assessed in the revalorization of groundwater quality. The groundwater quality was assessed on the basis of physical and chemical analysis (electric conductivity, pH, total dissolved solids (TDS), total hardness, NO3-, NO2-, SO4-2, Fe, Mn, Zn, Cu, B, residual sodium absorption (RSC) and sodium absorption ratio (SAR) for the period 2000-2004. From the analysis of results, it emerges that there are significant differences on the quality of water among the sample areas studied. The degradation of groundwater quality is mainly due to the pollution caused by the rural use of land, as well as its intensive exploitation. The salination and toxicity are potential problems of groundwater quality, especially in some areas, indicating that there is a need to take direct actions for the purpose of the optimum management of water resources in the Region of Thessaly. © 2007 Elsevier B.V. All rights reserved

Groundwater quality and location of productive activities in the region of Thessaly (Greece)

In the present study the involvement of human activities is assessed in the revalorization of groundwater quality. The groundwater quality was assessed on the basis of physical and chemical analysis (electric conductivity, pH, total dissolved solids (TDS), total hardness, NO3-, NO2-, SO4-2, Fe, Mn, Zn, Cu, B, residual sodium absorption (RSC) and sodium absorption ratio (SAR) for the period 2000-2004. From the analysis of results, it emerges that there are significant differences on the quality of water among the sample areas studied. The degradation of groundwater quality is mainly due to the pollution caused by the rural use of land, as well as its intensive exploitation. The salination and toxicity are potential problems of groundwater quality, especially in some areas, indicating that there is a need to take direct actions for the purpose of the optimum management of water resources in the Region of Thessaly. © 2007 Elsevier B.V. All rights reserved

Molecular characterization of a new intergenotype Norovirus GII recombinant

Human noroviruses (NoVs) of the Caliciviridae family are a major cause of epidemic gastroenteritis. The NoV genus is genetically diverse and recombination of viral RNA is known to depend upon various immunological and intracellular constraints that may allow the emergence of viable recombinants. In the present study, we report the development of a broadly reactive RT-PCR assay, which allowed the characterization of strain A6 at molecular level, established its genetic relationship at the sub-genogroup level and classified A6 strain at the sub-genotype level. The detection was carried out initially by enzyme-linked immunosorbent assay (ELISA) and the subsequent detection and molecular characterization of NoV strain was achieved by reverse transcription-PCR and sequencing. Based on the sequence analysis, A6 strain was revealed to belong to the GII genogroup of NoVs. Partial ORF1 gene sequencing analysis and complete ORF2 gene sequencing revealed that ORF1 and ORF2 belonged to two distinct genotypes GII/9 and GII/6, respectively, making obvious that A6 strain is a rare intergenotypic recombinant within the genogroup GII between GII.9 and GII.6 genotypes. A6 strain represents the first human NoV from Greece, whose genome has been partially (ORF1&ORF3) and completed (ORF2) sequenced. To our knowledge the recombination event GII.9/GII.6 in RdRp and capsid gene, respectively, that was revealed in the present study is reported for the first time

Influenza virus genotypes circulating in central greece during 2012-2014 and vaccine strain match

During the period 2012-2014 in central Greece, influenza A(H1N1), A(H3N2) and B virus genotypes were detected and isolated from individuals with influenza illness. Influenza A(H1N1) and A (H3N2) viruses were the dominant virus type in circulation as it was detected in 9% and 12% of pharyngeal swabs examined by real-time RT-PCR during both seasons, respectively, while type B viruses were detected in only 3% of the samples examined. Influenza activity in central Greece, as was determined by number of reported influenza cases and influenza positive samples detected, was markedly increased in the 2013-14, as compared to the 2012-13 season. Influenza A(H1N1)pdm09 viruses were detected during the 2013-14 season along with A (H3N2) and type B viruses. All type A(H1N1), A (H3N2) and type B influenza isolates analyzed by cell culture reacted to a high titer (> 640) against antisera to vaccine-like viruses of the same period, indicating satisfactory influenza vaccine protection against circulating seasonal and pandemic influenza viral strains. © 2015, Internet Scientific Publications, LLC. All rights reserved