Quantitative Detection Of HBV DNA Research Articles

Quantitative analysis of hepatitis B virus DNA based on raman spectroscopy combined with multivariate statistical methods

Quantitation of hepatitis B virus deoxyribonucleic acid (HBV DNA) is the most accurate method for determining the infection level of hepatitis B. It is a very meaningful means of detecting the different replication states of HBV infection. In this study, HBV DNA was detected and quantified in 499 patients with hepatitis B by confocal Raman spectroscopy combined with multivariate analysis. The effective information related to HBV DNA in spectral data was extracted by different pretreatments according to the vibrational modes of different functional groups. Quantitative analysis of HBV DNA was carried out by using the spectrum in the characteristic range of 807.027–1700.29 cm−1 for spectral peak assignment analysis and using partial least squares to construct a quantitative model. The results show that the regression model is established by combining normalization pretreatment and multivariate analysis methods. The correlation coefficient (Rc) and root mean square error of calibration from the calibration set of the selected feature range are 0.9751 and 0.3881, and the correlation coefficient (Rp) of the verification set and the root mean square error of prediction from the prediction set are 0.9673 and 0.4314. respectively. In clinical medical applications, Raman spectroscopy combined with diversified analysis methods has great research potential for the quantitative detection of HBV DNA in HBV serum.

Prevalence of HBV DNA among 20 million seronegative blood donations in China from 2010 to 2015.

The prevalence of HBV DNA among seronegative blood donations in China has not been studied extensively on a nationwide scale. The aim of this study was to analyse the prevalence, trend, distributions, and serological characteristics of HBV DNA positive/seronegative blood donations. We collected HBV test data from all blood banks of China from 2010 to 2015 and performed supplemental serological tests and quantitative detection of HBV DNA of the seronegative/HBV DNA positive blood donations. We analysed the prevalence of HBV DNA among seronegative blood donations screened by varying nucleotide acid test (NAT) reagents. The analysis results showed that a total of 20,084,187 seronegative blood donations were screened by NAT from 2010 to 2015 in China. The average frequency of HBV DNA among seronegative blood donations was 1/1482, but there has been a steady increase from 1/1861 in 2011 to 1/1269 in 2015. The geographical distribution of seronegative and HBV DNA positive blood donations was roughly consistent with that of HBsAg. The most common serological pattern was HBeAb and HBcAb positive. In conclusion, our study offeres fundamental data of seronegative and HBV DNA positive blood donations throughout China.

Epidemiologic Survey of Blood Donors with HBsAg⁻/HBV DNA⁺ in Chinese Xiamen Area

To understand the characteristics of infections from blood donors with HBsAg⁻/HBV DNA⁺ in Xiamen area. Donors in Xiamen area were assayed by routine ELISA and those with negative results were tested by nucleic acid amplification testing (NAT). HBsAg⁻/HBV DNA⁺ samples were tested by quantitative detection of HBV DNA. Epidemiological analysis and following up examination were conducted in HBsAg⁻/HBV DNA⁺ donors. Out of 130659 samples 113 were tested as HBsAg⁻/HBV DNA⁺ and with a rate of 0.09%. Among those, 62 samples were tested by quantitative detection of HBV DNA. All of the quantitative results were less than 1 × 10³ IU/ml and 93.5% (58/62) of which were less than 100 IU/ml. The possitive rate of HBsAg⁻/HBV DNA⁺ donors rose with ages. The possitive rate in male donors was higher than that in female and was lower in highly educated ones. Students and public servants had a lower positive rate. The possitive rate of HBsAg⁻/HBV DNA⁺ donors is higher in Xiamen and the distribution of possitive donors has certain epidemiological characteristics. It is necessary to mobilize and recruit more people with a lower rate of HBsAg⁻/HBV DNA⁺ infection.

Clinical, laboratory, and virological characteristics of patients with positive hepatitis B surface antigen in Upper Egypt

Hepatitis B virus (HBV) infection is a major cause of morbidity and mortality, particularly in developing countries. The aim of the study was to determine the clinical, laboratory, and virological characteristics of patients with chronic HBV infection in Upper Egypt. This descriptive, cross-sectional study included 252 patients with positive hepatitis B surface antigen (HBsAg). It was conducted in the Tropical Medicine and Gastroenterology Department and Outpatient Clinic, Assiut University Hospital (Egypt), from May 2012 to May 2014. All patients underwent clinical evaluation, were administered a questionnaire about risk factors for transmission of HBV, underwent liver function tests, abdominal ultrasonographic examination, and complete blood count, evaluation of serological markers of HBV, and quantitative detection of HBV-DNA. Of the 252 patients included, 88.5% were male with a mean age of 35.4 years. Arthralgia was the most common complaint (15.5%) and hepatomegaly was the most common finding (8.3%). As regards imaging results (ultrasonographic) the following were found: normal liver in 83.3%, coarse liver in 11.9%, hepatomegaly in 7.5%, splenomegaly in 6.3%, and cirrhosis in 5.9%. As regards laboratory results normal alanine aminotransferase was found in 79.8%, normal aspartate aminotransferase in 85.7%, reduced serum albumin in 4.4%, and low platelet count in 9.9%. The majority of patients (91.7%) were hepatitis B envelope antigen negative; 65.9% of patients were positive for HBV on PCR. No significant differences were found between positive HBV-DNA status (by PCR) and negative HBV-DNA status as regards clinical, imaging, and laboratory characteristics of patients. Most of the patients had normal liver on ultrasonographic examination and normal liver function tests. No significant difference was found between positive HBV-DNA status (by PCR) and negative HBV-DNA status as regards clinical, imaging, and laboratory characteristics of patients.

The clinical significance of quantitative detection of HBV DNA in the chronic infected patients

In recent years, antiviral therapy for chronic hepatitis B infected patients has achieved great development along with the invention of nucleos(t)ide, nucleos(t)ide analogues, interferon α and pegylated interferon α.The development of antiviral medicine also proposes new demands for the clinical diagnosis and therefore promotes the development of laboratory diagnostic techniques in the detection of chronic hepatitis B.In this review, we focused on the clinical application in the quantitive detection of HBV DNA, and its significance on clinical evaluation, treatment options, follow-up and prognosis in antiviral therapy.(Chin J Lab Med, 2012，35：117-121)

Microfabricated disposable DNA sensor for detection of hepatitis B virus DNA

Abstract The authors report here a preliminary evaluation of a microfabricated disposable-type DNA sensor, based on photolithography technique, for the detection of hepatitis B virus (HBV) genome DNA. The DNA sensor was reacted with the HBV-DNA-inserted plasmid (pYRB259), immersed in a 100 μmol/l Hoechst 33258 solution and washed with a phosphate buffer. The electrochemical analysis showed that the anodic peak current derived from Hoechst 33258 bound to the formed hybrids on the electrode was increased with increasing the concentration of pYRB259 in the range from 10 4 to 10 6 copy/ml. Then, the authors evaluated the concentration of HBV-DNA extracted from patients' sera using a DNA sensor and competitive polymerase chain reaction (C-PCR). A correlation coefficient of 0.89 was obtained between the two assay methods. The variance for each sample in triplicate measurements ranged from 2 to 6% in the inter-assay using the DNA sensor. These results suggest that the microfabricated disposable DNA sensor can provide a specific and quantitative detection of HBV-DNA in sera.

Quantitative detection of HBV DNA in serum using chemiluminescent revelation: Comparison with radioactive solution hybridization assay

Quantitative detection of HBV DNA in serum using chemiluminescent revelation: Comparison with radioactive solution hybridization assay

Advantage of PCR for detecting low amounts of HBV DNA in patients' sera

Serum hepatitis B virus DNA (HBV DNA) is now the most important and reliable marker for monitoring viral replication. Quantitative detection of HBV DNA in serum is based on a commercial standardized solution hybridization assay (Genostics). In this work, we studied the sensitivity and specificity of this method, in comparison with the polymerase chain reaction (PCR) technique, for low-value HBV DNA serum samples. Fifty-four patients with or without HBV serological markers were divided into 4 groups according to their HBV DNA values. Genomic amplication was found to affect 2 conserved regions of the viral genome, the S and C regions. Samples with an HBV DNA concentration equal to or greater than 1.5 pg/ml were considered positive in the “Genostics” test. A total of 38% of patients considered negative in the quantitative assay (< 1.5 pg/ml) were found to be positive for HBV DNA in serum after PCR. Only 26% of patients with an HBV DNA concentration of between 1.5 and 10 pg/ml in the Genostics test had PCR-detectable viral DNA in serum. Some 56% of patients with HBV DNA values between 10 and 20 pg/ml were found to be positive after amplification. All patients whose HBV DNA values were above 20 pg/ml had PCR-detectable viral DNA in serum. Our PCR results suggest that the positive limit level of the Genostics test has to be re-evaluated. Indeed, for low values of HBV DNA (under 20 pg/ml and especially under 10 pg/ml), it is not possible to conclude about the positivity from the quantitative assay, and results have to be estimated according to the clinical and serological status of the patients. Moreover, PCR can be falsely negative because of methodological problems. Nevertheless, this study confirms that PCR does enable detection of the viral genome in HBV-seronegative patients and in “old” and “cured” HBV-infection marker carriers.