Autoantibody immunoassays are critical in the diagnosis, disease stratification and prognosis of autoimmune disease (AID). Many important autoantibodies have been included in the diagnostic criteria for the classification and stratification of AID. However, the poor comparability among different methodologies may lead to significant differences in the interpretation of autoantibody test results, which poses a great challenge for the clinical application of autoantibody testing. The aim of this article is to discuss the main factors affecting the comparability of autoantibody test results, including the heterogeneity of autoantibodies, the variability of antigens, the setting of the positive judgment value for qualitative tests, and the traceability of calibrators for quantitative tests. In addition, this article also discusses the role of external quality assessment in improving the comparability of autoantibody test results.

Objective:To construct a method for hepatitis B virus (HBV) DNA detection based on recombinase-mediated isothermal amplification (RAA)-clustered regularly interspaced short palindromic repeats and their associated protein 13a (CRISPR-Cas13a).Methods:Through the alignment and screening of HBV DNA sequences, a positive plasmid was constructed, and recombinase-aided amplification (RAA) primers and CRISPR RNA (crRNA) were designed. A method for detecting HBV DNA based on the RAA-CRISPR-Cas13a system was developed, and the specificity and sensitivity were evaluated. Utilizing the CRISPR-Cas13a system, 70 clinical samples from HBV DNA-positive patients with various viral loads collected at Beijing You′an Hospital from 2019 to 2021 were analyzed. The detection results were further compared with those results using real-time quantitative polymerase chain reaction (qPCR).Results:The optimal RAA amplification primers and crRNA were first screened using the RAA-CRISPR-Cas13a method, with the sensitivities for detecting HBV DNA standards and for clinical samples at 1 IU/ml and<10 IU/ml, respectively, demonstrating specificity for HBV DNA detection. Compared with qPCR (the gold standard), the detection consistency between the two methods was 100% (70/70).Conclusion:This study established a method for detecting HBV DNA by integrating recombinase-aided amplification (RAA) technology with CRISPR/Cas13a technology.

Objective:Through the analysis of the detection results of hepatitis B virus (HBV) serological markers in the population admitted to Shengjing Hospital of China Medical University in recent five years, the prevalence and characteristics of HBV infection in our hospital were clarified.Methods:Cross-sectional study was conducted. A total of 1 017 030 patients who underwent HBV serological markers testing in Shengjing Hospital of China Medical University from January 1, 2019 to December 31, 2023 were enrolled as the research objects. The included cases were divided into 14 districts in Liaoning: Shenyang (345 346), Dalian (13 219), Anshan (38 536), Fushun (40 067), Benxi (28 883), Dandong (34 284), Jinzhou (35 827), Yingkou (40 573), Fuxin (30 675), Liaoyang (26 282), Panjin (21 008), Tieling (74 632), Chaoyang (43 858) and Huludao (20 949). The included cases were divided into 7 groups according to age: 0-10 years old (162 457), 11-20 years old (33 657), 21-30 years old (129 791), 31-40 years old (235 378), 41-50 years old (124 925), 51-60 years old (143 361), and≥61 years old (187 461). According to different time points of the implementation of vaccination policies, cases born from January 1, 1992 to December 31, 2023 were divided into three groups: the group from January 1, 1992 to December 31, 2001 (94 194), the group from January 1, 2002 to May 31, 2005 (70 428), and the group from June 1, 2005 to December 31, 2023 (87 057). The general data and HBV serological markers results were collected for statistical analysis. The comparison of intergroup rates was conducted using the Chi-square test, and the trend test was conducted using the Cochran Armitage test. Results:In recent five years, the total positivity of serum hepatitis B surface antigen (HBsAg), hepatitis B surface antibody (HBsAb), hepatitis B e antigen (HBeAg), hepatitis B e antibody (HBeAb) and hepatitis B core antibody (HBcAb) were 6.68% (67 938/1 017 030), 51.69% (485 627/939 498), 2.37% (21 961/926 626), 15.00% (138 853/925 759) and 27.86% (257 344/923 703), respectively. According to gender, the total positivity of HBV serological markers in men was significantly higher than that in women, and the difference was statistically significant (HBsAg: χ 2=5 599.286, P<0.001; HBsAb: χ 2=5.065, P=0.024; HBeAg: χ 2=2 451.420, P<0.001; HBeAb: χ 2=1 066.145, P<0.001; HBcAb: χ 2=4 013.618, P<0.001). According to the age group, the peak of HBsAg positivity was mainly distributed in the age group of 41-50 years old, with a positivity of 10.48% (2 864/27 324). The positivity of HBsAb decreased with age.The positivity of HBsAb decreased with age increase, highest in the 0-10 years old group, and a decrese in the 11-20 years old group ( Z=18 915.453, P<0.001). The positivity of HBcAb increased with age ( Z=27 493.853, P<0.001). According to the regional groups, the positivity of HBsAg and HBeAg in Shenyang City were 5.01% (3 996/76 974) and 1.45% (944/69 117) respectively. The positivity of HBsAb in patients from Liaoyang district was the highest [55.89% (3 362/6 150)]. The highest positivity of HBeAb and HBcAb were in patients from Dandong district, 19.90% (3 362/6 150) and 37.37% (3 119/8 680) respectively. According to the different time points of the implementation of the vaccination policies, the positivity of HBsAg in cases born from June 1, 2005 and December 31, 2023 decreased to 0.30% (259/87 057) compared with cases born before this time point (χ 2=5 777.47, P<0.001). Conclusions:The analysis of HBV serological laboratory characteristics showed that the positivity of contagious HBV serological markers was still high. The positivity of male was higher than that of female. HBsAb levels decreased significantly in the 11-20 years old group, suggesting the necessity to strengthen hepatitis B prevention in this age group. There were regional differences in the prevalence of hepatitis B, suggesting the necessity to optimize and improve HBV prevention and control strategies.

Objective:To explore the genetic diversity of Mycobacterium tuberculosis (MTB) in different types of tuberculosis and its association with clinical features, providing evidence for precise diagnosis and treatment of tuberculosis. Methods:This cross-sectional study included 103 cases of tuberculosis (38 with simple pulmonary tuberculosis, 43 with tuberculous pleurisy, and 22 with pulmonary combined with extrapulmonary tuberculosis) from Shenzhen Third People′s Hospital from 2015 to 2018. Paired bacterial strains from lung and pleural effusion/extrapulmonary sites were collected. Whole-genome sequencing (WGS) was used for drug resistance prediction, and genetic diversity (π value) was calculated as well as differential genes screening. Statistical analysis included paired t-tests and χ2 tests to compare clinical, bacteriological and genetic diversity features among groups.Results:The simple pulmonary tuberculosis group exhibited significantly higher rates of retreatment (71.7%, 27/38), cavitation (70.4%, 19/27), and multidrug-resistant or rifampicin-resistant (MDR/RR) (60.5%, 23/38) compared to the tuberculous pleurisy group (retreatment 11.9%, 5/42; cavitation 11.9%, 5/42; MDR/RR 16.3%, 7/43) and extrapulmonary tuberculosis group (retreatment 9.1%, 2/22; cavitation 18.2%, 4/22; MDR/RR 13.6%, 3/22) ( P<0.05). The overall π values of the MTB strain genomes in lung [(5.94±3.93)×10 ?5], pleural effusion[(6.22±3.51)×10 ?5], and extrapulmonary tissues [(5.83±3.54)×10 ?5] showed no significant differences ( H=0.10, P=0.94). Differential gene diversity analysis revealed that π value alternating genes related to respiration and intermediate metabolism were prominently high [tuberculous pleurisy 32.4% (11/34) and extrapulmonary tuberculosis groups 31.4% (32/102)], while cell wall-associated genes dominated in the simple pulmonary tuberculosis group (42.9%, 6/14). Drug resistance profiles and mutation spectra were identical across isolates from different sites within the same patient. Conclusion:WGS revealed the MTB diversity among different types of tuberculosis. Difference between pulmonary and extrapulmonary environments may impel the adaptive alternations of the bacterial strains to maintain survival with higher overall genome stability. Drug resistance testing of lung-derived isolates may provide references on extrapulmonary tuberculosis treatment.

The patient is a 60 year old male who has been experiencing thrombocytosis for over 2 years. He was treated outside the hospital in August 2023 due to acute myocardial infarction. After he left the hospital, there was still dizziness and persistent elevation of hemoglobin and platelets during follow-up. In November 2023, the patient went to Zhongshan Hospital affiliated with Fudan University for treatment. After admission, elevated platelets and hemoglobin were found, and genetic testing showed positive mutations in JAK2-V617F and CALR gene L367fs * 46, diagnosed as chronic myeloproliferative neoplasms (polycythemia vera and primary thrombocythemia). Discharge order was 0.1 g of hydroxyurea once a day. Follow up after six months showed improvement in hemoglobin and platelets compared with that at admission. This case suggests that classic driver gene mutation testing of myeloproliferative neoplasm is an important indicator for disease diagnosis for patients with thrombocytosis and hemoglobin increase.

Viral hepatitis and related liver diseases are among the most significant global healthcare issues. Viral hepatitis not only directly affects liver function but also interacts with the gut microbiota through the gut-liver axis. Imbalance in the gut microbiota may be associated with the occurrence, development and prognosis of viral hepatitis and related liver diseases. Detection and analysis of the gut microbiota can help to comprehensively understand the health status of patients with viral hepatitis and related liver diseases, providing a basis for early diagnosis. Recovery of gut microbiota dysbiosis may help to alleviate liver inflammation and to improve liver function. Regulating the gut microbiota through gut microbiota transplantation, probiotics and prebiotics may be new strategies for treating viral hepatitis and related liver diseases. By analyzing the latest research progress of gut microbiota in viral hepatitis and related liver diseases, this article provides new methods for the diagnosis and treatment of viral hepatitis and related liver diseases.

Immune thrombotic thrombocytopenic purpura (iTTP) is a fatal thrombotic microvascular disease closely associated with a severe deficiency of a disintegrin and metalloproteinase with thrombospondin motifs 13 (ADAMTS13). This review summarized the role of ADAMTS13 in the pathogenesis and clinical diagnosis and treatment of iTTP, as well as the detection of plasma ADAMTS13.

Many hematological disease prediction models built by big data analysis in medical laboratory and artificial intelligence (AI) technology, especially machine learning (ML) methods are reported more and more widely in researches on disease alerting, screening, diagnosis, discrimination, treatment and prediction. Because of a lack of guidance of standards, guidelines and normative documents in standardizing the construction and validation of these models, there is deficiency in model function, data preparation, screening and labeling, algorithm selection, model evaluation, as well as local and overall interpretation after model construction, which also limit the clinical application. Therefore, with integration of the literature review, combining the practice on blood disease prediction models, this article proposes that some keypoints and efficiency should be noticed, in order to build hematological diseases related models with clinical practical application value.

Objective:To identify the ABO blood group in a family with ABO forward/reverse typing discrepancies using serological methods and third-generation sequencing technology.Methods:In January 2024, samples with ABO blood group forward/reverse typing discrepancies were referred to Dalian Blood Center for blood group identification. Standard serological techniques were used to identify the ABO blood group. For the samples of the proband with a serological phenotype of AB weak and his father, full-length haplotype sequencing of the ABO gene was performed using third-generation sequencing. The impact of amino acid mutations on protein structure was also predicted. Results:Sequencing revealed that, compared to the reference sequence ABO*B.01, both the proband and his father had a haplotype with a mutation c.278C>T in Exon 7, resulting in p.Pro93Leu. This is consistent with the ABO*BW.12 genotype. The spatial structure of the ABO*BW.12 protein was altered, leading to reduced stability and impaired function of the B glycosyltransferase (GTB).Conclusion:For the identification of AB weak blood group, the combination of serological methods and third-generation sequencing technology enables accurate and efficient detection of haplotype mutations and identification of ABO subtypes, thus ensuring clinical transfusion safety.

Objective:To preliminarily establish a sample clot warning model for coagulation screening tests using 5 machine learning methods.Methods:This cross-sectional study collected 7 401 routine screening test samples from Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, from January 1st, 2015, to August 18th, 2024, including 4 786 clotted (positive) and 2 615 qualified (negative) samples for model development. The dataset was divided into Dataset 1 and Dataset 2 based on a reagent change for APTT in December 2018, with separate models developed for each. An additional 2 493 samples (October 31st to November 8th, 2024) were used to evaluate consistency between the model and manual assessment, while 23 200 samples (October 17th to December 31st, 2024) were used for assessing real-world predictive performance. Five machine learning algorithms were employed to develop the clot prediction model: logistic regression (LR), random forest (RF), extreme gradient boosting (XGBoost), naive bayes (NB), and artificial neural network (ANN), with the ANN model constructed using two different hidden layer and neuron parameter settings. Model selection was based on AUC, accuracy, sensitivity, specificity, F1-score, PPV, and NPV, with the optimal model integrated into the LIS for validation.Results:Among the six models using 5 machine learning algorithms, XGBoost demonstrated the highest performance (AUC=0.961, sensitivity=0.945, F1-score=0.934) and robustness to reagent changes ( Z=-1.333, P=0.113). When deployed, the differences between the model's predictions and manual pre-judgment were statistically significant ( Z=-5.289 to 8.933, all P<0.01). The predictive efficacy indices AUC (95% CI), sensitivity, specificity, and accuracy of the XGBoost model deployed in real-world operation of the LIS were 0.939 (0.918—0.960), 0.958, 0.921, and 0.921 respectively. Conclusion:In this study, a clot warning model for coagulation screening samples was established based on the XGBoost algorithm, and its prediction efficacy is good, providing a foundation for intelligent pre-analytical quality control for coagulation screening tests.