Analyze Risk Factors About Hemolytic Disease Of The Newborn And Verify Experimental Conditions Of Relevant Test

ObjectiveThrough reviewing literature, we want to screen the relevant risk factors of hemolytic disease of the newborn (HDN). Combined with clinical practice, aimed to solve the most common but debatable issue about HDN laboratory diagnostic tests-IgM antibody can affect the IgG antibody titer results. The experiment conditions of IgG antibody titer were researched, to increas the accurancy of laboratory diagnosis.Method1. Using HDN related search terms to complete literature search of Chinese and English document database. The HDN risk factors associated literatures were screened by reading literature titles and abstracts. The risk factors that may lead to HDN were statisticed.2.4 groups analog serum of difference titers were configured and the titers were distributed as follows:128+512,128+128,128+32,128+8 (monoclonal IgG+monoclonal IgM). The IgM antibodies were damaged by 2-Me at room temperature and 37℃ for 30min. The IgM and IgG antibody titers were detected by hemagglutination test and gel anti-human globulin test respectively, and the test should be completed before and after the IgM antibodies were destructed.3. Equal volume of 2-Me and pregnant plasma were incubated for 30min at room and 37℃temperature to destroy IgM antibodies. The IgM and IgG antibody titers were checked by saline hemagglutination and gel anti-human globulin test respectively. The test should be completed before and after the IgM antibodies were destructed.4. The IgG and IgM immunoglobulin qualitative were tested by immune protein electrophoresis and quantitative were detected by special protein analyzer, including before and after IgM antibodies were destroyed, to evaluate the effect of 2-Me treated IgM and IgG.Results1. The parent’s age, blood group incompatible, irregular antibodiy, antibody titer, number of pregnancies, abortion history, stillbirth and transfusion, and so on, may be related to HDN pathogenesis.2. The effect of 2-Me destroyed IgM antibodies at room and 37℃ temprature were no significant difference in 4 analog serum groups. When IgM≤IgG titers, IgM antibody can’t interfer IgG antibody titers, and the detection results were consistent with the real results. When IgM>IgG titers, IgM titers can mask IgG results, leading to IgG inaccurate results. In the 4 groups, IgG titers score have positive correlation with IgM titers. When IgM were destroyed, the 4 groups IgG titers score were no significant difference.3. After IgM antibodies were damaged, the IgM titers were significantly decline in human serum samples. The IgG antibody titers were no difference before and after IgM antibody damaged. After IgM were destructed, detecting IgG antibody titers scores were lower than undestroyed (p<0.05).4. Both patient and analog plasma samples were destroyed by 2-Me, there were increased in IgM Immunoglobulin quantitatively results. No matter whether the plasma was damaged, IgG and IgM qualitative detection were positive by immuno electrophoresis technology.Conclusion1. The parent’s age, blood group incompatible, irregular antibodies, antibody titers, number of pregnancies, history of abortion, stillbirth and transfusion, and so on, may be related to HDN pathogenesis.2. The IgM antibodies can be destroyed at room temperature for 30min, which can meet requirement of detecting IgG antibody titer by micro-gel method. In patients samples, when IgM≤IgG titers, IgM can’t affect the accuracy of IgG antibody titer detection.