ABSTRACT
ObjectiveThe traditional detection method of recombinase polymerase amplificaton (RPA) is not suitable for rapid field detection due to the complicated operation and other factors. Taking the detection of hepatitis b virus (HBV) nucleic acid as an example, it established a detection method of HBV nucleic acid isothermal amplification based on recombinase polymerase amplificaton (RPA) and designed a matching visual detection device of RPA product.MethodsFirstly, a RPA product detection device was designed, which can be used to collect images by taking photos of mobile phones and visually interpret the detection results. Secondly, RPA primers and probes were designed according to the design of HBV gene conserved sequence. Amplification efficiency of each primer pairs were compared though monitoring the RPA reaction of real-time fluorescence curve to screen the best primers and optimize the optimal reaction conditions. Visual detection sensitivity was investigated by using artificial synthesis of HBV target plasmid, and was investigated the specificity of the method by the detection of synthetic plasmid containing hepatitis c virus (HCV), human immunodeficiency virus, treponema pallidum, influenza virus, human papilloma virus DNA fragment. Thirdly, the feasibility of RPA product visualization detection device was verified by comparing with the real-time fluorescence amplification curve. Finally, RPA visual detection was performed on 20 serum DNA samples detected by real-time fluorescence PCR to evaluate the applicability of this method to the detection of actual clinical samples.ResultsThe visual detection device of RPA product was used to realize the negative and positive signals that could be detected by mobile phone photography and visual observation. The visual detection method of HBV nucleic acid RPA amplification could realize the visual detection of DNA targets as low as 1-10 copies of HBV within 30 min at 39 ℃ and had good specificity. The test results of 20 serum DNA samples were completely consistent with those of the commercially available qPCR kit, which preliminarily verified the practicability of the method and the device.ConclusionCombined the established HBV-RPA amplification system with the RPA product visualized detection device, it would be expected to develop a low-cost rapid visualization screening technology platform for HBV nucleic acid in blood.
ABSTRACT
This study attempts to discuss the correlation between UGT1A1*28 as uridine diphosphate glucuronosyltransferase gene promoter and coding region Gly71Arg gene polymorphism with neonatal hyperbilirubinemia of neonates in Wuhan.A total of 168 neonates were divided into the hyperbilirubinemia group (case group,n=108) and healthy neonates group (control group,n=60).Their DNA was obtained through blood extraction.The gene exon mutation of UGT1A1 was detected by Sanger sequencing,which revealed the relationship between UGT 1A 1*28 and Gly71Arg polymorphism with neonatal hyperbilirubinemia of neonates.The results showed that:(1) The frequency of UGT1Al*28 allele mutation in the case group and the control group was 9.3％ and 10％ respectively,with the difference being not significant between the two groups (P＞0.05).(2) The frequency of Gly71Arg allele mutation in the case group and the control group was 35.1％ and 21.7％ respectively,with the difference being significant between the two groups (P＜0.01).(3) The serum bilirubin level of Gly71Arg mutant homozygous and heterozygous subgroups (n=66) in the case group was 302.7±31.4 μmol/L,which was significantly higher than 267.3±28.5 μmol/L of the wild subgroup (n=42) (P＜0.01).It was suggested that the occurrence of neonatal hyperbilirubinemia of neonates in Wuhan was not associated with UGT 1A1*28 gene polymorphism,but closely with the Gly71Arg gene polymorphism.Meanwhile,the Arg allele mutation was related to the degree of jaundice.
ABSTRACT
<p><b>OBJECTIVE</b>To evaluate the cost-effectiveness of combining Chinese medicine (CM) with Western medicine (WM) for ischemic stroke patients.</p><p><b>METHODS</b>Hospitalization summary reports between 2006 and 2010 from eight hospitals in Beijing were used to analyze the length of stay (LOS), cost per stay (CPS), and outcomes at discharge.</p><p><b>RESULTS</b>Among 12,009 patients (female, 36.44%; mean age, 69.98±13.06 years old), a substantial number of patients were treated by the WM_Chinese patent medicine (CPM)_Chinese herbal medicine (CHM) (38.90%); followed by the WM_CPM (32.55%), the WM (24.26%), and the WM_CHM (4.15%). With adjustment for confounding variables, LOS of the WM_CPM_CHM group was about 10 days longer than that of the WM group, and about 6 days longer than that of the WM_CPM group or the WM_CHM group (P<0.01); CPS of the WM_CPM_CHM group was United States dollar (USD) 1,288 more than that of the WM group, and about USD 600 more than that of the WM_CPM group or the WM_CHM group (P<0.01). Compared with the WM group, odd ratio (OR) of recovered and improved outcome of the WM_CPM_CHM group was the highest [OR: 12.76, 95% confidence intervals (CI): 9.23, 17.64, P<0.01], OR of death outcome of the WM_CPM_CHM group was the lowest (OR: 0.08, 95% CI: 0.05, 0.12, P<0.01). There was no significant difference between LOS, CPS and OR of the WM_CPM group and those of the WM_CHM group (P>0.05). Cost/effectiveness and incremental cost-effectiveness ratio of the WM_CPM_CHM group were robustly higher than those of the WM group.</p><p><b>CONCLUSION</b>Compared with WM alone, supplementing CPM and CHM to WM provides significant health benefits of improving the chance of recovered and improved outcome, and reducing the death rate, at an expense of longer LOS and higher CPS.</p>