ABSTRACT
AIMS: To investigate the effects of heparin in detection of LIAISON® Rubella IgM (Rub-M) and the mechanism of interference. METHODS: Different concentrations of lithium heparin and sodium heparin were added to ten serum samples. The relative light units (RLU) value of Rub-M was measured using the LIAISON XL detection system. Different levels of IgM serum were incubated with magnetic particle in Rub-M detection kit at 4 °C for 4 h, blocking anti-human IgM-specific antibodies coated on the surface of magnetic particle. Separately, the rubella virus antigen in Rub-M detection kit was replaced by phosphate-buffered saline (PBS). The RLU values of LIAISON® Rub-M of original serum and serum containing various concentrations of heparin were measured after the above two different treatments. RESULTS: The RLU value of LIAISON® Rub-M increased with the increase of heparin content lower than 40 IU/mL, and reached a peak value at 40-50 IU/mL. The RLU value of LIAISON® Rub-M then decreased with the decrease of heparin concentration. When rubella virus antigen was replaced by PBS, the RLU value of LIAISON® Rub-M of serum samples containing 40 IU/mL heparin decreased significantly. The blocking concentration of IgM increased gradually, and the RLU value of LIAISON® Rub-M of seven serum samples containing 40 IU/mL heparin also decreased gradually. CONCLUSION: Plasma with heparin cannot be used to the detection of LIAISON® Rub-M. Heparin may participate in the reaction by binding with rubella virus antigen and anti-human IgM-specific antibodies coated on the surface of magnetic particle, thus affecting the detection results.
Subject(s)
Heparin , Rubella , Antibodies, Viral , Humans , Immunoglobulin M , Rubella/diagnosis , Rubella virusABSTRACT
Polymalic acid (PMA) is a novel biopolymer produced by the polyextremotolerant fungus Aureobasidium pullulans. In this study, a GATA-family transcriptional factor, Gat1, which regulates nitrogen uptake and PMA biosynthesis, was investigated. PMA production increased to 11.2% in the mutant overexpressing gat1 but decreased to 49.1% of the PMA titre when gat1 was knocked out from the genome of A. pullulans. Comparative transcriptome analysis of wild-type and mutant strains (∆gat1 and OE::gat1) revealed that 23 common differentially expressed genes were related to oxidative phosphorylation, ribosome biogenesis, and nitrogen metabolism. Under nitrogen-limited stress, regardless of the preferred nitrogen (glutamine, Gln) or non-preferred nitrogen (proline, Pro), 70% of Gat1 in the cells was located in the nucleus-cytoplasm, which resulted in an increase in nitrogen uptake and PMA biosynthesis regulation. Quantitative RT-PCR revealed that glucosekinase (GLK) in the glycolytic pathway and malate synthase (MLS) in the glyoxylate shunt pathway may be cross-regulated by Gat1 and nitrogen concentration (Gln or Pro), Therefore, glk was overexpressed in mutant strain (OE::gat1), which resulted in an increased PMA titre and yield of 12.6% and 13.0% respectively. These findings indicate that Gat1 may play an important role in the dual regulation of the nitrogen and carbon metabolisms in PMA biosynthesis.
