Comparative study of anti-double stranded DNA detection by ELISA and Crithidia luciliae immunofluorescence

Two commercial enzyme-linked immunosorbent assays (ELISA) and two commercial Crithidia luciliae immunofluorescence tests (CLIF) were reevaluated as to the efficiency and degree of correlation of anti-double stranded DNA (anti-dsDNA) detection in systemic lupus erythematosus (SLE). The two ELISAs exhibited an overall agreement of 95% and significantly correlated with each other (r=0.91, p<0.001). They were comparable in sensitivity (64%, 61%) and had the same specificity (95%, 95%). The sensitivity of the two CLIFs was 39% and 29% with corresponding specificities of 100% and 97%, and an overall agreement with each other of 94%. The two ELISAs had comparable specificity to the CLIFs with good agreement (84%, 79%) while they had a much greater sensitivity than the CLIFs. These findings suggest that ELISA is a useful laboratory test for anti-dsDNA detection of SLE due to its simplicity, quantitative results, sensitivity, specificity and cost, as compared to CLIFs.     

Visual detection of white spot syndrome virus using DNA-functionalized gold nanoparticles as probes combined with loop-mediated isothermal amplification

The integration of loop-mediated isothermal amplification (LAMP) and DNA-functionalized AuNPs as visual detection probes (LAMP–AuNPs) was developed and applied for the detection of white spot syndrome virus (WSSV) from Penaeid shrimp in this study. The principle of this combination assay relies on the basis of stability characteristics of the DNA-functionalized AuNPs upon hybridization with the complementary target DNA toward salt-induced aggregation. If the detected target DNA is not complementary to the ssDNA probes, the DNA-functionalized AuNPs will be aggregated due to the screening effect of salt, resulting in the change of solution color from red to blue/gray and shift of the surface plasmon peak to longer wavelength. While the DNA-functionalized AuNPs are perfectly matched to the detected target DNA, the color of solution still remains red in color and no surface plasmon spectral shift. This assay provides simply technique, time-saving and its detection results could be achieved qualitatively and quantitatively by visualization using the naked eye due to the colorimetric change and by measurement using the UV–vis spectroscopy due to the surface plasmon spectral shift, respectively. In this study, LAMP–AuNPs assay was successfully developed with the detection of WSSV-LAMP generated product at 0.03 μg/reaction, and showed the sensitivity of 2 × 10² copies WSSV plasmid DNA, that is comparable to the most sensitive method reported to date. The LAMP–AuNPs assay described in this study revealed a highly sensitive, rapid and reliable diagnostic protocol for detection of WSSV. This technique has a potential as a routine method for assessing the infectious diseases in Penaeid shrimp not only for WSSV, but also for other shrimp pathogens, and can be useful tool in field conditions for the diagnosis or surveillance programs.     

Changes in prevalence, awareness, treatment and control of hypertension in Thai population, 2004-2009: Thai National Health Examination Survey III-IV

OBJECTIVE:: To determine the changes in prevalence, awareness, treatment and control of hypertension and their metabolic risk factors in Thai population between 2004 and 2009. METHODS:: The Thai National Health Examination Survey (NHES) in 2004 and 2009 data were used. Blood pressure and anthropometric measurements were performed. Prevalence, awareness, treatment and control of hypertension of Thai population aged at least 15 years were calculated. Analyses were weighted to the probability of sampling. RESULTS:: The prevalence of hypertension in 2004 and 2009 were relatively stable at approximately 21.0%. There was improvement in awareness of hypertension, from 18.2% for men and 33.0% for women in 2004 to 39.5 and 59.4% in 2009, respectively. The high blood pressure control rates improved from 4.8 to 14.4% for men and from 10.8 to 27.2% for women, respectively (all P?相似文献   

Five-year prospective study of tuberculin skin testing among new healthcare personnel at a university hospital in Thailand

We determined the prevalence of a positive tuberculin skin test (TST) and the incidence of TST conversion among new healthcare personnel (HCP) in a hospital in Thailand. During 2005-2008, TST was performed on 1438 HCP and the prevalence of positive TST was 66.3%. Age, male gender, and the presence of Bacille Calmette-Guérin (BCG) scar were associated with odds of positive TST (all P < 0.05). The incidence of TST conversion was 4.8 per 100 HCP-years. Nine (0.6%) HCP were diagnosed with active tuberculosis. The annual surveillance programme is important for the early diagnosis and prevention of tuberculosis among HCP in Thailand.     

Detection of anti-dsDNA by ELISA using different sources of antigens

AIM: To compare different sources of DNA for use in ELISA-based assays for anti-dsDNA antibody detection in systemic lupus erythematosus (SLE) diagnosis. METHOD: Bacterial genomic DNA from Flavobacterium menignosepticum, Proteus vulgalis, Seratia marcescens, Streptococcus pyogenes and Salmonella typhimurium and genomic DNA from human blood were used as antigens for IgG anti-dsDNA detection by enzyme-linked immunosorbent assay (ELISA). Eighty-six sera were tested, 28 derived from patients with SLE, 28 from patients with other rheumatic diseases and 30 from normal human subjects. RESULTS: Genomic DNA from Flavobacterium menignosepticum and human blood had high sensitivity (75%, 82%) and specificity (91%, 91%) for anti-dsDNA detection in diagnosis of SLE. However, human genomic DNA was the most effective antigen of all antigens studied. The assay had a higher sensitivity but lower specificity than commercial ELISA (61% sensitivity and 95% specificity). There was a high level of correlation between commercial ELISA and ELISA using human genomic DNA as antigen (r=0.776, p<0.001) and they exhibited a high level of diagnostic agreement with each other (kappa=0.890, p<0.001). CONCLUSION: The genomic DNA from human blood is a potentially useful source of antigen for the detection of anti-dsDNA by ELISA. However, further studies are required to compare the performance of ELISA using this source of antigen against commercial radioimmunoassays for anti-dsDNA detection.     

Prevalence and management of diabetes and metabolic risk factors in Thai adults: the Thai National Health Examination Survey IV, 2009

OBJECTIVE

To determine the prevalence of impaired fasting glucose (IFG) and undiagnosed and diagnosed diabetes in Thai adults in 2009 and examine the extent of changes in proportions of diagnosis, treatment, and control for blood glucose, high blood pressure, and high total cholesterol between 2004 and 2009.

RESEARCH DESIGN AND METHODS

Data from the multistage cross-sectional National Health Examination Survey (NHES) IV of 18,629 Thai adults aged ≥20 years conducted in 2009 were used to analyze and compare with the data from NHES III in 2004.

RESULTS

The prevalence of IFG and diabetes was 10.6 and 7.5%, respectively. Of all diabetes diagnoses, 35.4% were not previously diagnosed, and the proportion was higher in men than in women (47.3 vs. 23.4%, P < 0.05). Compared with those in year 2004, the proportions of individuals with diabetes and concomitant hypertension did not significantly decrease in 2009 in both sexes, but the proportions of women with diabetes who were abdominally obese or had high total cholesterol (≥5.2 mmol/L) significantly increased in 2009 by 18.0 and 23.5%, respectively (all P < 0.01). The rates of treatment and control of blood glucose, high blood pressure, and high total cholesterol were favorably improved in 2009. However, in substantial proportions of individuals with diabetes these concomitants were still controlled suboptimally.

CONCLUSIONS

The prevalence of diabetes and IFG remained high in Thai adults. Improvement in detection and control of diabetes and associated metabolic risk factors, particularly obesity and high serum cholesterol, are necessary.Diabetes has become a major global public health burden. It has been estimated that the number of people with diabetes worldwide was 285 million in 2010 and will increase to 439 million in 2030, with the majority of increase (69%) occurring in developing countries (1). The estimated number of individuals with diabetes in Asia was 113 million in 2010 and will increase to 180 million in 2030 (2). The increase has been the result of the rise in obesity as a consequence of changes in lifestyle toward urbanization, with high energy intake and low physical activity following the rapid economic growth and urbanization in this region (2).In Thailand, a low-middle income country, diabetes has been a major cause of morbidity and mortality in the past decade (3). Diabetes alone is responsible for 3.3 and 8.3% of total deaths in Thai men and women, respectively (3). A high prevalence rate of diabetes in Thailand makes it among the top ten in Asia (2). In 2004, the National Health Examination Survey (NHES) III reported a prevalence of 6.7% in adults aged ≥15 years, of whom 53.3% went undiagnosed. The prevalence of impaired fasting glucose (IFG) is 12.5% (4). Undiagnosed diabetes increases the risk of complications as a result of being untreated, and about 40% of those treated have their fasting plasma glucose (FPG) under control (<7.8 mmol/L) (4).To monitor the diabetes situation, the fourth NHES was conducted in 2009. This study aimed to determine the prevalence of IFG, diabetes, and associated metabolic risk factors in Thai adults aged ≥20 years and older in 2009. Among individuals with diabetes, we also examined the extent of changes in proportions of metabolic risk factors and lack of diagnosis, treatment, and control for high blood glucose, high blood pressure, and high total cholesterol between the years 2004 and 2009.