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- Evander, Magnus, et al.
(författare)
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Puumala hantavirus viremia diagnosed by real-time reverse transcriptase PCR using samples from patients with hemorrhagic fever and renal syndrome
- 2007
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Ingår i: Journal of Clinical Microbiology. - : American Society for Microbiology. - 0095-1137 .- 1098-660X. ; 45:8, s. 2491-2497
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Tidskriftsartikel (refereegranskat)abstract
- Puumala virus (PUUV) is the endemic hantavirus in northern Sweden and causes nephropathia epidemica (NE), a milder form of hemorrhagic fever with renal syndrome. There is a need for fast and reliable diagnostics to differentiate the disease from other infections. By aligning virus RNA sequences isolated from 11 different bank voles and one human patient, we designed a real-time reverse transcriptase (RT) PCR method for detection of PUUV RNA. The real-time RT-PCR assay showed linearity from 20 to 2 x 10(6) virus copies with a correlation coefficient above 0.98 to 0.99 for all experiments. The detection threshold for PUUV cDNA was two copies per reaction. A two-step qualitative RT-PCR to detect PUUV RNA showed 100% concordance with the real-time RT-PCR assay. PUUV RNA viremia was detected in 33 of 34 PUUV immunoglobulin M (IgM)-positive patients with typical clinical NE disease from the region of endemicity. One PUUV IgM-negative sample had PUUV RNA, and 4 days later, the patient was IgM positive. Of samples with indeterminate IgM, 43% were PUUV RNA positive. The kinetics of antibody titers and PUUV viremia were studied, and five of six NE patients displayed a decrease in PUUV viremia a few days after disease outbreak coupled with an increase in PUUV IgM and IgG. In one patient with continuously high PUUV RNA levels but low IgM and no IgG response, the infection was lethal. These findings demonstrated that real-time RT-PCR is a useful method for diagnosis of PUUV viremia and for detecting PUUV RNA at early time points, before the appearance of IgM antibodies.
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- Karefylakis, Christos, 1982-, et al.
(författare)
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Vitamin D C3 epimer in a mid-Swedish region : Analytical measurement and epidemiology
- 2018
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Ingår i: Clinica Chimica Acta. - Amsterdam, Netherlands : Elsevier. - 0009-8981 .- 1873-3492. ; 478, s. 182-187
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Tidskriftsartikel (refereegranskat)abstract
- Background: The discovery of an epimeric form of 25(OH)D3 may complicate the interpretation of vitamin Dstatus. The aim of this study was to examine the prevalence and determinants of 25-hydroxy-3-epi-vitamin D3 (3- epi-25(OH)D3) in a mid-Swedish region and to investigate how the measurement of 3-epi-25(OH)D3 would affect the assessment of vitamin D status using current thresholds.Methods: We conducted a cross-sectional study of 8286 in- and outpatients in primary as well as secondary care settings. Plasma 25(OH)D, 25(OH)D2, 25(OH)D3 and 3-epi-25(OH)D3 were measured using High Pressure Liquid Chromatography – Tandem Mass Spectrometry (HPLC – MS/MS). The relative 3-epi-25(OH)D3 contribution was calculated as a percentage of the total 25(OH)D3. Blood samples were collected between March 2014 and July 2015 providing a seasonal aspect to the results.Results: 3-epi-25(OH)D3was detected in 635 cases (7.7% of all subjects), and the mean concentration was8.4 ± 3.5 nmol/L. 3-epi-25(OH)D3correlated significantly with 25(OH)D3(r =0.38, p < 0.001).A multivariateanalysis among the detected showed that male gender and winter season were independently associatedwith higher 3-epi-25(OH)D3/25(OH)D3percentage ratio (R2=0.044). Infants and children had a significantlyhigher detection rate compared to the reference age category (18–45 years) as well as those who were testedduring the summer season.Conclusions: We report findings from the first epidemiologic study of 3-epi-25(OH)D3 conducted in Sweden, based on a large population sample. 3-epi-25(OH)D3 was detected in 7.7% of the study population and the mean concentration was 8.4 nmol/L. The quantification of 3-epi-25(OH)D3 would not significantly influence the clinical interpretation of vitamin D levels. Additional studies are needed to understand the metabolic pathway and the possible physiological functions of this metabolite.
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