ABSTRACT
Objective:To investigate the specificity and sensitivity of four methods including ultrafastreal-time fluorescence polymerase chain reaction (PCR), real-time fluorescence (RT)-PCR, enzyme-linked immunosorbent assay (ELISA) and gold immunochromatography assay (GICA) for the detection of novel Bunya virus, so as to provide experimental basis for the early diagnosis of severe fever with thrombocytopenia syndrome (SFTS).Methods:Serum samples from 86 clinically diagnosis SFTS patients admitted to the Jinan Infectious Diseases Hospital Affiliated to Shandoug University were tested by ultrafast real-time fluorescence PCR, RT-PCR, ELISA and GICA during June 1 to September 30, 2017. Chi-square test was used for statistical analysis.Results:Among 86 serum samples, the positive rate of novel Bunya virus of ultrafast real-time fluorescence PCR, RT-PCR, IgM-ELISA, IgG-ELISA, IgM-GICA and IgG-GICA were 82(95.34%), 79(91.86%), 41(47.67%), 8(9.3%), 19(22.09%) and 3(3.49%), respectively. The specificity of ultrafast real-time fluorescence PCR was 100%, and the sensitivity was 1×10 3 copies/mL.Repeated amplification test showed that the variation coefficient of the computed tomography value was <2%.During phases one, two and three, the positive rates of ultrafast real-time fluorescence PCR were 41(97.62%), 34(94.44%) and 7(87.50%), and RT-PCR were 39(92.86%), 33(91.67%) and 7(87.50%), respectively. During phases one and two, the positive rate of ultrafast real-time fluorescence PCR was slightly higher. The positive rate of anti-novel Bunya virus antibody (IgM) tested by ELISA had a significant increase from phase one (28.57%)to phase three (87.50%). There were statistical differences between phase two and phase, as well as between phase three and phase one ( χ2=8.347 and 7.561, respectively, both P<0.01). IgM-GICA also had an increase from phase one (14.29%) to phase two (33.33%)( χ2=3.962, P<0.01), while it was still lower than the other tests.In phase one, the positive rate of RT-PCR was higher than those of ELISA(both IgM and IgG)and GICA(both IgM and IgG)( χ2=33.740, 55.080, 49.010 and 64.340, respectively, all P<0.01). In phase two, the positive rate of RT-PCR was higher than those of ELISA(both IgM and IgG)and GICA(both IgM and IgG) ( χ2=7.700, 46.720, 23.700 and 50.630, respectively, all P<0.01). In phase three, the positive rates of ultrafast real-time fluorescence PCR, RT-PCR and IgM-ELISA were equivalent, which were all higher than those of IgG-ELISA and GICA (both IgM and IgG). The positive rates of RT-PCR and IgG-ELISA, IgM-GICA and IgG-GICA were significantly different (all χ2=6.250, all P<0.05). Conclusion:In the early detection of novel Bunya virus, ultrafast real-time fluorescence PCR has higher sensitivity, specificity, good repeatability and high stability, which greatly reduces the amplification time compared with the traditional RT-PCR, and is of great value in the early and rapid diagnosis of SFTS.
ABSTRACT
BACKGROUND: Umbilical cord mesenchymal stem cells (UC-MSCs) have made certain curative effect on hepatic failure, but little is reported on the effect of hepatic failure patient's serum microenvironment on UC-MSCs differentiation ability. OBJECTIVE: To investigate the effect of hepatic failure patient's serum on the differentiation of umbilical cord mesenchymal stem cells into hepatic cells. METHODS: UC-MSCs were isolated by tissue adherent method and the cell morphology and phenotype identified by microscope and flow cytometry. Alpha-MEM media with serum before/after plasmapheresis were used to culture the hirdgeneration of UC-MSCs, and regular fetal bovine serum culture medium acted as control group. Inverted microscope was used to observe the cell morphology in three groups. Immunohistochemical method was used to measure expression level of alpha fetoprotein, albumin and cytokeratin 18. RESULTS AND CONCLUSION: A great amount of high-purity UC-MSCs could be obtained using tissue adherent method, which highly expressed CD44, CD73, CD90, CD10, but did not express CD45. Hepatic failure patient's serum could change the morphology of UC-MSCs and induce UC-MSCs to express alpha fetoprotein, albumin and cytokeratin 18. The positive expression of alpha fetoprotein, albumin and cytokeratin 18 was significantly increased after plasmapheresis (P < 0.05). To conclude, hepatic failure patient's serum after plasmapheresis exert more benefits to induce UC-MSCs to differentiate into hepatic cells than that before plasmapheresis.
ABSTRACT
Objective To provide data evidence for brucellosis prevention by a retrospective analysis of brucellosis epidemiologic feature in Ji'nan Infectious Disease Hospital from 2010-2014.Method The brucellosis patients with confinned diagnosis from 2010 to 2014 admitted to Ji'nan Infectious Disease Hospital were included in this study,and their epidemic data were statistically analyzed,including time,age and occupation distributions.Results There were 1 010 brucellosis patients in total in Ji'nan Infectious Disease Hospital from 2010 to 2014,and 12 cases,69 cases,134 cases,274 cases and 521 cases,respectively,in each year,which showed rapid rising trend.There were two peak times,the first was from April to August,the incidence was 9.1％ (92/1 010),12.2％(123/1010),13.4％ (135/1010),11.1％ (112/1010) and 10.7％ (108/1010),respectively,in each month;the second was January,the incidence was 9.4％ (95/1010).The diagnosed rate within 1 week of onset only accounted for 13.4％ (135/1010),and 72.4％ (731/1010) patients were diagnosed for more than two weeks after the onset.The male incidence of brucellosis was higher than the female,and the sex ratio was 2.36:1.00.In terms of age distribution,40-49 and 50-59 were both peak ages,with incidence of 29.3％ (296/1 010) and 25.0％ (252/1010),respectively.The professional was given priority to farmers and herdsmen (85.6％,865/1 010),and 96.4％ (974/1010)of the patients had a clear exposure history,mainly related to sheep,cattle,pigs and other livestock,also related to pets and milk.Conclusions The brucellosis patients that received in Ji'nan Infectious Disease Hospital are increased significantly in the recent 5 years,and their transmission is closely associated with livestock breeding and processing.Most patients have failed timely diagnosis in the early days,which should catch enough of our attention.