ABSTRACT
Objective To evaluate the performance of magnetic nanoparticle chemiluminescence immunoassay (NM-CLIA) in detection of allergen-specific immunoglobulin E (sIgE) antibodies to Dermatophagoides pteronyssinus (International Allergen Code D1) and Dermatophagoides farinae (International Allergen Code D2). Methods A total of 489 serum samples from the patients with suspected allergic disease (244 cases caused by D1, and 245 caused by D2), who were treated at Suzhou Kowloon Hospital, Shanghai Jiaotong University School of Medicine, were detected by NM-CLIA and immunofluorescence assay, respectively. χ2 test and Kappa test were used to evaluate the correlation between the two methods in detection of D1 and D2 sIgE antibodies. The limit of detection (LoD), linear range and precision of NM-CLIA in detection of D1 and D2 sIgE antibodies were verified by the standard method of American Clinical Laboratory Standardization Association. Results The LoDs of NM-CLIA in detecting D1 and D2 sIgE antibodies were both less than 0.01 U/mL, the linearity ranged from 0.1 to 100 U/mL, the within-run precision was less than 5%, and the between-run precision was less than 8%. Methodological comparison results showed that NM-CLIA and immunofluorescence assay had good consistency in detecting D1 and D2 sIgE antibodies. For D1, the positive coincidence rate and negative coincidence rate were 95% and 92%, respectively (χ2＝174.45, P0.001, Kappa＝0.843), and the ±1 class agreement was 95.6%; for D2, the positive coincidence rate and negative coincidence rate were 91% and 97%, respectively (χ2＝154.263，P0.001，Kappa＝0.787), and the ±1 class agreement was 94.2%. Conclusion NM-CLIA has good correlation with immunofluorescence assay in detecting D1 and D2 sIgE antibodies, and has good LoD, linear range and precision, suggesting that it can be recommended for clinical testing of D1 and D2 sIgE antibodies.
ABSTRACT
Because of limited viral replication and lack of cytopathic effect in cell culture, a new PCR-based rapid seroneutralization assay for detection of GII.4 norovirus neutralized antibodies was developed with serum samples from acute-phase patients, convalescent-phase patients and healthy controls. According to this study, neutralizing antibodies were detected in 100% of convalescent-phase sera, and in 2.5% of healthy controls sera. However, all of the acute-phase serum samples could not neutralize virus efficiently. Compared to the results from ELISA (96.2% at sensitivity and 80% at specificity), the present in vitro neutralization assay is more specific and more sensitive.
Subject(s)
Humans , Antibodies, Neutralizing , Allergy and Immunology , Base Sequence , Caliciviridae Infections , Diagnosis , Virology , Case-Control Studies , Cell Line , DNA Primers , Enzyme-Linked Immunosorbent Assay , Gastroenteritis , Diagnosis , Virology , Norovirus , Allergy and Immunology , Polymerase Chain Reaction , Methods , Sensitivity and SpecificityABSTRACT
<p><b>OBJECTIVE</b>To determine the relationship between polymorphisms in the genes encoding IL-1, IL-6, and IL-10 with primary biliary cirrhosis (PBC) in Chinese population.</p><p><b>METHODS</b>Whole-blood samples were taken from 77 patients with PBC and 160 healthy controls. DNA was extracted and the polymorphisms at positions IL-1 +3953, IL-1RN intron 2, IL-6 -174, and IL-10 -1082, -819, and -592 were determined by using sequence-specific polymerase chain reaction (SSP) or polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP).</p><p><b>RESULTS</b>The frequency of IL-1RN1,1 allele in PBC group was significantly higher than in control group (90.9% vs 79.4%, P=0.026), and the frequency of IL-1RN1,2 in PBC group was significantly lower than in control group (6.5% vs 18.8%, P=0.013). There was no significant difference in the frequence of IL-1RN*2 allele between PBC group and control group (P=0.06). Of the 77 patients with PBC, 4 patients were IL-6 -174GC, 73 were IL-6 174GG. All the 160 health controls are IL-6 -174GG (P=0.0036). The frequence of IL-6 -174C allele in PBC group was significantly higher than that in control group (P=0.0038). No significant differences of polymorphisms for IL-1 +3953 and IL-10 (-1082, -819 and -592) were found between PBC group and control group.</p><p><b>CONCLUSION</b>The polymorphisms of IL-1RN and IL-6 -174G/C appear to be associated with PBC, and the polymorphisms of IL-1 +3953 and IL-10 promoter gene are not associated with PBC in a Chinese population.</p>