Objective: Hepatitis B virus (HBV) has a worldwide distribution and remains a leading public health problem in China. Method: Automated chemiluminescence microparticle immunoassay was used to test all five markers of HBV serology in serum samples among 696,048 patients, pregnant women, and normal subjects in Beijing from 2008 to 2018. Results: The overall prevalence of subjects categorized as previous/ occult HBV infection, inactive HBsAg carrier, active HBV infection, HBsAg, HBV susceptible, and immune via vaccination was 29.4%, 4.8%, 1.4%, 6.4%, 33.9% and 30.3%, respectively; men had a significantly higher prevalence of HBV infection than women. The prevalence of HBsAg was around 0.5% in subjects <= 10 years of age, increased dramatically to 3.7% in subjects between 11 and 20 years of age, reached the highest level of 7.9% in subjects between 41 and 50 years of age, and finally decreased to 2.8% in subjects >= 81 years of age. During the 10 years from 2008 to 2018, the prevalence of HBsAg was stabilized at about 6.0%, and indicators of HBV susceptibility, previous/ occult HBV infection, and immunity via vaccination were not further improved, despite the constant implementation of HBV vaccination since 1992. All four age groups (21 - 30y, 31 - 40y, 41 - 50y and 51 - 60y) of the normal adult population were found to have a significantly lower prevalence of HBsAg and HBV susceptibility but significantly higher prevalence of immunity via vaccination compared with corresponding age groups of the sub-total population. Conclusions: Although high coverage has been established among infants and young children, their vaccination alone could not reduce HBV infection in the adult Chinese population quickly. Adult populations with more vaccinated individuals are found to have fewer individuals with HBsAg. Vaccination in adults or at least in high-risk adults is an urgent need to decrease horizontal HBV transmission in China. (C) 2020 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY-NC-NDlicense (http://creativecommons.org/licenses/by-nc-nd/4.0/).

BackgroundPrimary cytomegalovirus (CMV) infection is prevalent worldwide and usually results in latency in immunocompetent populations. Reactivation of latent CMV can cause life-threatening complications in immunocompromised hosts.MethodsWe used the CMV Brite assay to test CMV antigenemia (pp65) in whole blood samples from 22,192 patients with or without autoimmune diseases in Beijing during 2008-2018.ResultsThe overall prevalence of CMV antigenemia was 19.5% (9.7%, males; 26.0%, females). The prevalence of CMV antigenemia was 35.1%, 58.6% and 11.4% in whole patients with autoimmune diseases, in patients with systemic lupus erythematosus (SLE) and in patients with non-SLE autoimmune diseases, respectively. All patients with non-autoimmune diseases, patients with HIV/AIDS or transplantation were found to have 5.0%, 27% or 14.8%, respectively. Patients≤20 years with SLE had a significantly higher prevalence of CMV antigenemia than did all SLE patients, on average. Patients>51 years with non-SLE autoimmune diseases had a significantly higher prevalence than did all patients with non-SLE autoimmune diseases, on average. The prevalence of CMV antigenemia in patients admitted to intensive-care units (ICUs) were 9.2%, which was significantly higher than that among all patients with non-autoimmune diseases. Patients with SLE had 23.8% of negative conversion of CMV antigenemia, significantly lower than the percentage of patients with non-SLE autoimmune (64.3%) and non-autoimmune (61.0%) diseases. The mean number of days to negative conversion of CMV antigenemia in patients with SLE was 35.3±35.8 days, which was significantly longer than that in patients with non-SLE autoimmune diseases (15.4±11.9 days) and non-autoimmune diseases (13.6±7.7 days).ConclusionsCMV antigenemia is found more likely in women than in men, more prevalently in patients with SLE than those with HIV/AIDS or transplant recipients, more frequently in patients admitted to ICUs. Patients with SLE had prolonged CMV antigenemia. The role of CMV appears important in SLE.

Background Chlamydia pneumoniae (C. pneumoniae) is an obligate intracellular bacterium and a human pathogen that causes respiratory infectious diseases. More than 50% of the adult population worldwide was once infected with C. pneumoniae, but investigations into this topic are insufficient in mainland China. Methods Anti-C. pneumoniae IgG and IgM antibodies were detected using micro-immunofluorescence test in serum samples of patients visiting Peking Union Medical College Hospital between 2008 and 2017 for routine medical purposes, and the aim of this retrospective study was to analyze the test results. Results Among 12,050 serum specimens tested for anti-C. pneumoniae IgG and IgM antibodies, the overall prevalence of anti-C. pneumoniae IgG antibodies was 86.6%, 87.2% for men and 86.0% for women. Adult men (>20 years) were found to have a significantly higher prevalence of anti-C. pneumoniae IgG than women (χ2 = 30.32, P = 0.000). 3 to 5 years old patients were observed to have the lowest prevalence of anti-C. pneumoniae IgG, 42.8%, then increased with age, reaching the highest level of 98.6% in patients over 70 years of age. In the 10,434 specimens with C. pneumoniae IgG antibodies, the total geometric mean titer (GMT) for C. pneumoniae IgG was 45.71. Although GMTs were found to be significantly higher among all men than among all women (t = 5.916, P = 0.000), sex difference actually began in patients over 40 years of age and increased in the elderly. In the total 12,050 specimens, 1.2% had anti-C. pneumoniae IgM, 3.3% had anti-C. pneumoniae IgG with titers equal to or greater than 1:512; 0.39% had ≥4-fold increasing titers of antibodies in acute and convalescent phase paired samples, and 4.4% were finally confirmed to have acute antibodies against C. pneumoniae. 6 to 10 years old patients were found to have the highest rate of both IgM antibodies (3.9%) and acute antibodies (6.2%) against C. pneumoniae. Acute antibodies against C. pneumoniae were found to be more frequent in patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD, 14.0%, χ2 = 20.43, P = 0.000), patients with pneumonia (7.8%, χ2 = 51.87, P = 0.000) and patients with acute respiratory tract infection (12.3%, χ2 = 60.91, P = 0.000) than among all patients (4.4%). Both anti-C. pneumoniae IgG and IgM antibodies should be tested for acute antibodies against C. pneumoniae as testing for either alone will underestimate by a maximum of two-thirds the incidence of acute antibodies against C. pneumoniae. Conclusions More than 86% of Chinese patients on an average were once infected with C. pneumoniae. Adult men had both a higher prevalence and higher levels of antibodies than women. 6 to10 year old patients were found to have the most frequent acute infection of C. pneumoniae. C. pneumoniae is associated with AECOPD, pneumonia and acute respiratory tract infection. Anti-C. pneumoniae IgG and IgM should be tested simultaneously to avoid underestimation of acute antibodies against C. pneumoniae.

IntroductionDuring routine blood measurements using an automated hematology analyzer, two easily confused types of suspect flags related to lymphocytes often appear: atypical and immature lymphocytes. The aim of this study was to investigate the correlation of high fluorescence cell (HFC) parameter and lymphocyte flags determined from an automated hematology analyzer.MethodsA total of 93 patients affected by various pathologic conditions (viral infection, immunological disease, oncological disease and tumor) were divided into an “atypical lymphocytes” group (“atypical” for short), an “immature lymphocytes/blasts” flag group (abnormal), a mixed‐flag group that includes “atypical lymphocytes” (mixed), and a non‐flag group (non‐flag).ResultsThe numbers of HFCs in the atypical, abnormal, mixed, and non‐flag groups were 1.8% (0.9%‐5.5%), 0.7% (0.1%‐5.0%), 2.3% (1.2%‐5.0%), and 0.8% (0.7%‐1.2%), respectively. The HFCs of “atypical” appeared as a separate cluster with clear boundaries. The HFCs of “abnormal” as an unclear boundaries, and it was difficult to accurately distinguish between the HFCs from the immature lymphocytes and the normal lymphocytes. The lower limit of HFC when the atypical lymphocyte flag appeared was 0.04 × 109/L. The number of HFCs was similar to atypical lymphocytes detected by microscopy and CD19+CD20−CD27++ cells by flow cytometry at 78% and 76%, respectively. The number of HFCs detected in “atypical” and CD19+CD20−CD27++ cells showed good consistency (r = .715), whereas the consistency was poorest for “abnormal” (r = .176).ConclusionIt demonstrates that HFCs reflects atypical lymphocytes better than immature lymphocytes/blasts.

Objective To prepare monoclonal antibody against early and late antigens of cytomegalovirus (CMV) and to use the antibody for the identification of isolates from proficiency testing and clinical samples.Methods BALB/C mice were immunized with soluble and formaldehyde to inactivate CMV antigens from the CMV-infected MRC-5 cells at 24,48 and 72 hours after inoculation.Mouse myeloma NS-1 cells were fused with spleen cells of the immunized mice.Positive hybridomas were screened with EIA and were cloned with limited dilution method.The immunofluorescent and immunoblotting tests were applied to confirm the positive clones after screening.The ascites was prepared with the identified hybridoma and was purified by Protein G.The purified monoclonal antibody was used to identify CMV isolates from CAP proficiency testing samples and clinical samples.Results A total of 110 positive hybridomas clones were preliminary identified from fused cells of three immunized mice.One hybridoma clone,23B5-1 was finally obtained while other clones were eliminated due to cross-reactive with other antigens,low antibody titers or without reactions with both CMV early and late antigens.It was shown that 23B5-1 reacted with CMV infected MRC-5 cells from 3 hours to 120 hours (5 days) after inoculation.All of immediate-early,early and the late CMV antigens were recognized by the monoclonal antibodies simultaneously.It was also shown that 23B5-1 reacted with five CMV protein bands between 25KD to 50 KD in western blot.The immunoglobulin subclass of antibody was IgG1 and the titer of the purified antibody was equal or more than 1:12 800.The results of staining with the antibody were consistence with predicted results for all six CMV isolates from proficiency testing and ten isolates from clinical samples.Conclusions The performance of our self-prepared monoclonal antibody was good according to the preliminary application.