Establishment and preliminary verification of a highly sensitive nucleic acid extraction method for OBI detection
Journal Title: Chinese Journal of Blood Transfusion - Year 2024, Vol 37, Issue 2
Abstract
Objective To establish and verify a new nucleic acid extraction method for OBI detection with large volume and high sensitivity, and apply it in the quantitative determination of OBI samples with low viral load. Methods The method for nucleic acid extraction with large volume was established based on the method of Roche nucleic acid detection kit. HBV standards were configured into 10 000 IU/mL, 1 000 IU/mL, 100 IU/mL, 10 IU/mL and 1 IU/mL respectively, and nucleic acid was extracted from the 10 mL standards by magnetic beads. CT values of each concentration were detected by fluorescence quantitative PCR and each concentration gradient was detected in parallel duplicates. The logarithm of virus concentration was taken as the X-axis and the average CT values of two tests were taken as the Y-axis to construct the fluorescence quantitative standard curve and regression equation. Three repeated experiments were conducted to verify the stability of the method. This method was used to extract nucleic acid from OBI samples with low viral load, and fluorescence quantification was performed. Results The amplification efficiency of fluorescence quantitative standard curves ranged from 90% to 105%, and the regression equation was greater than 0.99. The variation coefficients of variation of CT values were 0.63%, 0.78%, 1.52%, 1.36% and 0.78%, respectively. This method can extract nucleic acid from OBI samples with viral load of 1 IU/mL for quantification. Conclusion The detection limit of HBV nucleic acid quantitative detection system can reach 1 IU/mL, and it has strong stability and high sensitivity, which can be used for the quantitative detection of OBI with low viral load.
Authors and Affiliations
Wenbo GAO, Bo HE, Rongsong DU, Fenfang LIAO, Junmou XIE, Min WANG, Hao WANG
Keywords
Related Articles
- EP ID EP738764
- DOI 10.13303/j.cjbt.issn.1004549x.2024.02.010
- Views 74
- Downloads 0
Evaluation and analysis of bleeding risk of anticoagulation therapy in severe non-ICU patients with COVID-19 based on heparinase-modified TEG
Objective To investigate the clinical application of heparinase-modified TEG (hmTEG) in evaluating coagulation status and monitoring anticoagulant therapy in severe non-ICU patients with COVID-19. Methods The clinical da...
Molecular mechanism of ABO bood group variation
Objective To study the relationship between ABO subtype, para-Bombay blood group and genotype, so as to explore the possible molecular mechanism of these two blood groups, and provide accurate genetic detection targets a...
Impact of blood screening strategies for Hepatitis C virus on blood safety
Objective To establish a reasonable and effective blood screening strategy for Hepatitis C virus (HCV), so as to reduce the risk of blood transfusion transmission, ensure blood safety and improve the quality of blood scr...
Application effectiveness of PDCA in emergency blood management
Objective To explore the effectiveness of PDCA (Plan-Do-Check-Act Cycle Management) in clinical emergency blood management. Methods The data of emergency blood-using cases from January 2021 to June 2022 in each clinical...
IgA deficiency allergic transfusion reaction and its prevention and treatment strategies
IgA deficiency allergic transfusion reaction often occurs in patients with anti-IgA. When blood products containing IgA are transfused into these patients, it may lead to severe allergic transfusion reactions, which can...