Preparation of colloidal gold test strips for the detection of antibodies to peste des petits ruminants based on monoclonal antibodies to N protein / 生物工程学报

A simple, fast, and visual method for detecting antibodies against peste des petits ruminants virus (PPRV) using colloidal gold strips was developed. In this study, the pET-32a-N was transformed into Escherichia coli Rosetta (DE3) for expression. Hybridoma cell lines were generated by fusing SP2/0 myeloma cells with splenocytes from immunized mice with the expressed and purified N protein of PPRV. The PPRV N protein was labeled with colloidal gold particles as the gold-labeled antigen. The N protein served as the gold standard antigen and as the test (T) line-coated antigen, while the monoclonal antibody served as the quality control (C) line-coated antibody to assemble the colloidal gold immunochromatographic test strips for detecting antibodies against the N protein of PPRV. Hybridoma cell line designated as 1F1 was able to stably secrete the monoclonal antibody against the N protein of PPRV. The titer of 1F1 monoclonal antibody in ascites was 1:128 000 determined by indirect enzyme-linked immunosorbent assays (ELISA), and the immunoglobulin subtype of the monoclonal antibody was IgG1, with kappa chain. The obtained monoclonal antibody was able to specifically recognize the N protein of PPRV, as shown by Western blotting and indirect immunofluorescent assay (IFA). The developed colloidal gold test strip method was able to detect PPRV antibodies specifically, and there was no difference between different batches of the test strips. Testing of a total of 122 clinical sera showed that the compliance rate of the test strip with ELISA test was 97.6%.The test strip assay developed in this study has good specificity, reproducibility, and sensitivity, and it can be used for the rapid detection of PPRV antibodies.

Urinary immunoglobulins in viral diagnosis: An overview

Antibody detection by serological methods gained a lot of interest in recent years and has become the backbone of virological diagnosis. Despite the detection of all five classes of immunoglobulins in urine, not much attention has been paid to the use of urine as a diagnostic sample to detect viral antibodies. Unlike venipuncture, this non-invasive mode of sample collection can help cover all age groups, especially paediatric and old age patients, where blood collection is difficult. Using urine as a sample is also economical and involves lesser risk in sample collection. The antibodies are found to be stable in urine at room temperature for a prolonged period, which makes the sample transport management easier as well. A few recent studies, have also shown that the detection limit of antibodies in urine is at par with serum or other clinical material. So, the ease in sample collection, availability of samples in large quantity and stability of immunoglobulins in urine for prolonged periods can make urine an ideal sample for viral diagnosis.

Three platelet antibody detection methods: a comparative study / 中国输血杂志

【Objective】 To compare three kinds of platelet antibody detection methods used to identify alloantibodies in patients with platelet transfusion refractory(PTR). 【Methods】 The 83 samples from PTR patients were analyzed base on three different methods, including solid phase ELISA, Luminex, and capture. The sensitivity, reproducibility, and consistency of different kits were evaluated. 【Results】 A total of 71 (62 positive and 9 negative) out of 83 samples showed consistent results by three methods. The consistency between Luminex and solid phase ELISA was 95.2% (Kapp value=0.829, P<0.05), between solid phase ELISA and capture method was 85.5% (Kapp value=0.512, P<0.05), and between Luminex and capture method was 90.3% (kappa value=0.636, P<0.05). Among the 12 samples with inconsistent results, 3 cases presented positive results by capture method alone and negative by other methods, which had incompatible cross-matching results with 6 random blood donors; 5 cases with HLA antibodies showed negative results by capture method alone and positive by both Luminex and solid phase ELISA; the other 4 cases were positive in both capture and Luminex, but negative in solid phase ELISA. 【Conclusion】 The consistency of three methods was 85.5%, and each has its limitations. The capture method is rapid, economic and registered domestically, which can be used for preliminary screening.Luminex has the optimal diagnostic performance, which can be used for high-throughput and HPA/HLA antibody analysis. The solid phase ELISA is convenient. The combination of them can detect platelet antibodies effectively.

Establishment and clinical evaluation of dry fluorescent luminescencemethod for detection of antibodies to hepatitis C virus / 中国输血杂志

【Objective】 To establish a dry fluorescent luminescence method for the detection of antibodies to hepatitis C virus (HCV) and evaluate its clinical application. 【Methods】 Anti-HCV antibody was detected by double-antigen sandwich dry fluorescent luminescence method established using multi-epitope chimeric antigen. The established method was used to detect national reference samples(positive 20, negative 20), and a total of 349 clinical samples, including 108 HCV patients, 36 patients with other diseases and 205 healthy individuals, which were tested in parallel with enzyme-linked immunoassay (ELISA) to evaluate the performance of the established method. 【Results】 The concordance rate of positive and negative(each 20) reference samples were both 100% (20/20), and the CV of precision reference sample was 9.16%, which met the requirements of national reference samples. In clinical performance evaluation, the AUC value was 0.984, and the sensitivity and specificity of the dry fluorescent luminescence method were 96.30% (104/108) and 96.27% (233/241). The overall concordance rate between dry fluorescent luminescence method and ELISA was 97.71% (341/349) (Kappa=0.952). 【Conclusion】 The dry fluorescence luminescence method of HCV antibody is simple and rapid, with high sensitivity and high specificity, and can be used in clinical application.

Blood group serological testings of ABO hemolytic disease of newborn: A retrospective analysis / 中国输血杂志

【Objective】 To retrospectively analyze blood samples of suspected hemolytic disease of newborn(HDN)mothers and infants, and detect hemolytic disease caused by irregular antibodies, so as to provide help for the clinical diagnosis and treatment of HDN. 【Methods】 632 suspected HDN samples from Obstetrics and Pediatrics Department of our hospital from January 2016 to October 2018 were collected, and serologically detected by microcolumn gel technique, as well as DAT, free antibody test and antibody release test. 【Results】 Among 632 samples, 306 were HDN positive, with a positive rate at 48.4%, 64 suspected HDN, accounting for 10.1%, and 262 non confirmed HDN, accounting for 41.5%. 180 samples were type A, among which 145 were HDN positive, with a positive rate at 80.56%; 233 were type B, among which 157 were HDN positive, wiht a positive rate at 67.38%; 210 were type O, among which 1 was HDN positive, with a positive rate at 0.48%; 9 were type AB, among which 3 were HDN positive, with a positive rate at 33.33%. The positive rates of HDN differed by blood types (P<0.05). In 632 suspected HDN samples, 9 were with irregular antibody + immune anti-A, and 136 with solo immune anti-A; 10 were with irregular antibody + immune anti-B, and 170 with solo immune anti-B; 1 was with irregular antibody + immune anti-A and anti-B, and 2 with immune anti-A and anti-B; 4 HDN cases were caused by irregular antibody, while anti-S and anti-E cconstituted 2 and 2 cases, respectively. 【Conclusion】 ABO HDN is more common and attracts more attention in clinical practice than HDN scaused by other group systems, which were rare and easy to be ignored, but also may cause moderate and severe HDN. even severe anemia, edema and stillbirth of fetus. Therefore, it is necessary to carry out irregular antibody screening during pregnancy so as to achieve early detection and treatment.

Blood screening strategy for SARS-CoV-2 among low risk populations / 中国输血杂志

【Objective】 To analyze the viability of 2 different blood screening strategies against SARS-CoV-2 in low risk populations, so as to provide references for the formulation of blood screening strategy. 【Methods】 Two screening strategies for antibodies against SARS-CoV-2 were adopted: 1) the total antibody were initially screened for all samples, and the antibody IgG and IgM were retested in those primary positive samples; 2) only antibody test of IgG and IgM for all samples. And SARS-CoV-2 nucleic acid was detected in parallel. Reactive samples was confirmed by neutralization test. The sensitivity, specificity and true positive rate of two strategies were calculated. 【Results】 None was positive for SARS-CoV-2 nucleic acid among 880 samples. Four truly positive samples were implicated in 9 (1.02%, 9/880) initially reactive samples in total antibody test; 3 in 26 (2.95%, 26/880) initially IgG or IgM reactive samples. 【Conclusion】 The first strategy is superior to the second strategy in the sensitivity and specificity, and is recommended for the detection of SARS-CoV-2 antibody in low risk populations.

Preparation of different fragments of SARS-CoV-2 N protein and its application in fluorescence chromatography / 生物工程学报

Different fragments of SARS-CoV-2 nucleocapsid (N) protein were expressed and purified, and a fluorescence immunochromatography method for detection of SARS-CoV-2 total antibody was established. The effect of different protein fragments on the performance of the method was evaluated. The N protein sequence was analyzed by bioinformatics technology, expressed in prokaryotic cell and purified by metal ion affinity chromatography column. Different N protein fragments were prepared for comparison. EDC reaction was used to label fluorescence microsphere on the synthesized antigen to construct sandwich fluorescence chromatography antibody detection assay, and the performance was systemically evaluated. Among the 4 prepared N protein fragments, the full-length N protein (N419) was selected as the optimized coating antigen, N412 with 0.5 mol/L NaCl was used as the optimal combination; deleting 91-120 amino acids from the N-terminal of N412 reduced non-specific signal by 87.5%. the linear range of detection was 0.312-80 U/L, the limit of detection was 0.165 U/L, and the accuracy was more than 95%. A fluorescence immunochromatographic detection method for analysis of SARS-CoV-2 total antibody was established by pairing N protein fragments. The detection result achieved 98% concordance with the commercially available Guangzhou Wanfu test strip, which is expected to be used as a supplementary approach for detection of SARS-CoV-2. The assay could also provide experimental reference for improving the performance of COVID-19 antibody detection reagents.

Evaluation of the diagnostic efficiency of three anti-Echinococcus antibody-based assays for the serodiagnosis of echinococcosis / 中国血吸虫病防治杂志

ObjectiveTo evaluate the efficiency of three Chinese commercial anti-Echinococcus antibody-based assays for the serodiagnosis of echinococcosis. MethodsA total of 142 sera from cystic echinococcosis patients, 89 sera from alveolar echinococcosis and 39 sera from healthy controls were sampled, and detected by kits A (ELISA), B (ELISA) and C (colloidal gold immunoassay). The routine blood testing results and biochemical parameters were compared between the cystic and alveolar echinococcosis patients, and the associations of the absorbance (A value) of the serum specific antibody detected by A and B kits with the routine blood testing results and biochemical parameters were examined in echinococcosis patients. In addition, the performance of these three assays for the serodiagnosis of echinococcosis was evaluated. Results There were no significant differences between the cystic and alveolar echinococcosis patients in terms of the median white blood cell count (WBC), neutrophil count (NEU), monocyte count (MONO), basophil count (BASO), alanine aminotransferase concentration (ALT), aspirate aminotransferase concentration (AST), total bilirubin (TBIL), direct bilirubin (DBIL), indirect bilirubin (IBIL) (all P values > 0.05), and higher median lymphocyte count (LYM) and albumin levels (ALB) were detected in cystic echinococcosis patients than in alveolar echinococcosis patients (both P values < 0.05), while the median eosinophil count (EOS) was greater in the alveolar echinococcosis patients than in the cystic echinococcosis patients (P < 0.01). The A value of the serum specific antibody detected by kit A showed a linear positive correlation with WBC (rs = 0.153, P < 0.05) and EOS (rs = 0.174, P < 0.05), and a linear negative correlation with TBIL (rs = -0.134, P < 0.05) and IBIL (rs = -0.146, P < 0.05), while the A value of the serum specific antibody detected by kit B showed a linear positive correlation with WBC (rs = 0.257, P < 0.01), NEU (rs = 0.203, P < 0.01), MONO (rs = 0.159, P < 0.05), EOS (rs = 0.330, P < 0.01), ALT (rs = 0.171, P < 0.01) and AST (rs = 0.160, P < 0.05), and a linear negative correlation with ALB (rs = -0.168, P < 0.05). The overall coincidence rate, sensitivity, specificity, Youden’s index and Kappa value of A, B and C kits were 86.30%, 69.63% and 91.48%; 84.42%, 64.94% and 92.21%; 97.44%, 97.44% and 87.18%; 0.82, 0.62 and 0.79; and 0.600, 0.337 and 0.750 for the diagnosis of echinococcosis, respectively. The overall coincidence rate, sensitivity, specificity and Youden’s index of A, B and C kits were 84.54%, 64.64% and 71.82%; 80.99%, 55.63% and 68.31%; 97.44%, 97.44% and 87.18%; and 0.78, 0.53 and 0.56 for the diagnosis of cystic echinococcosis, respectively, while the overall coincidence rate, sensitivity, specificity and Youden’s index of A, B and C kits were 92.19%, 85.16% and 85.16%; 89.89%, 79.78% and 84.27%; 97.44%, 97.44% and 87.18%; and 0.87, 0.77 and 0.72 for the diagnosis of alveolar echinococcosis, respectively. The C kit showed cross-reactions in the serodiagnosis of cystic echinococcosis and alveolar echinococcosis. There were no significant difference in the area under the receiver operating characteristic curve (ROC) between A and B kits for the diagnosis of echinococcosis (0.970 vs. 0.948, Z = 1.618, P > 0.05), and there was a high agreement between A and B kits in the diagnosis of echinococcosis (Kappa = 0.585, P < 0.01). Conclusions The three commercial anti-Echinococcus antibody-based kits exhibit a higher serodiagnostic efficiency for alveolar echinococcosis than for cystic echinococcosis. The A kit shows a high sensitivity and specificity for the diagnosis of echinococcosis, and has a relatively stable diagnostic performance and fewer influencing factors, which is suitable for the pre-surgical preliminary diagnosis and post-surgical follow-up monitoring of serum anti-Echinococcus antibody, while the C kit shows a high sensitivity and specificity for the diagnosis of echinococcosis, and is easy to perform and high in reporting rate, which is feasible for initial screening of echinococcosis.

HIV/AIDS case detection and influencing factors for late diagnosisin Hangzhou from 2013 to 2018 / 预防医学

Objective@#To analyze current situation of HIV/AIDS case detection and factors associated with late diagnosis among the newly diagnosed cases from 2013 to 2018 in Hangzhou, so as to provide basis for improving the detection capacity of HIV. @*Methods@#The data of HIV testing and newly diagnosed HIV/AIDS cases in Hangzhou from 2013 to 2018 were collected through the China AIDS Prevention and Control Information System. The proportion of HIV antibody detection and positive cases in different regions, detection ways and high-risk groups were analyzed. The influencing factors for late diagnosis were analyzed by multivariate logistic regression model. @*Results@#The proportions of cases with HIV detected, HIV positive and late diagnosis increased from 2013 to 2018, and the annual ones were 24.99%, 6.95 per ten thousand and 30.07%, respectively. The results of the multivariate logistic regression analysis showed that people who were male ( OR=1.656, 95%CI: 1.351-2.030 ) and aged older ( OR: 1.912-5.117, 95%CI: 1.250-7.904 ) had higher risks of late diagnosis; who detected HIV through pre-test of receiving blood ( OR=4.429, 95%CI:2.217-9.225 ) , other inpatient detection ( OR=2.137, 95%CI: 1.615-2.826 ) , preoperative testing ( OR=2.137, 95%CI: 1.615-2.826 ) and testing of STD clinic attendants ( OR=1.359, 95%CI: 1.007-1.834 ) had higher risks of late diagnosis compared to those diagnosed at VCT clinics; who diagnosed at CDCs ( OR=0.714,95%CI: 0.558-0.915 ) and community health centers ( OR=0.645, 95%CI: 0.441-0.943 ) had lower risks of late diagnosis than those diagnosed in hospitals; who were infected by heterosexual contact ( OR=1.299, 95%CI: 1.130-1.493 ) had a higher risk of late diagnosis than MSM; who had history of STD ( OR=0.818, 95%CI: 0.706-0.948 ) had a lower risk of late diagnosis than who did not.@*Conclusions@# HIV testing and case detection had been expanded, but late diagnosis had not been improved in Hangzhou from 2013 to 2018. Age, sex, route and institution of diagnosis, transmission route and history of STD were influencing factors of late diagnosis.

Current status and research progress of serological antibody detection of Mycoplasma pneumonia / 国际儿科学杂志

Mycoplasma pneumonia(MP) is a common cause of children community-acquired pneumonia,and can lead to a variety of extrapulmonary complications.Thus,a rapid,sensitive diagnostic method is particularly important for early diagnosis and treatment.As MP grows slowly,requires additional nutrient supply and easy to be contaminated by fungi and bacteria during the culture process,culture of MP might not be suitable for clinical detection.However,the PCR technique is more complicated and high requirement for operator to be implemented in primary hospitals.Therefore,the serum diagnosis has become the commonly used method in clinical diagnosis.This paper is to review the latest findings of different serological diagnostic methods,antigen component,and antibody type in different periods of infection of MP.

Problems in serum antibody detection of smear-and culture-negative pulmonary tuberculosis / 中华微生物学和免疫学杂志

More than 60% of active tuberculosis(TB) patients are smear- and culture-negative, constituting a prime group in the prevention and control of TB in China. In the existing laboratory testing technologies, immunological diagnosis is more advantageous than etiological diagnosis in the detection of smear-and culture-negative TB. Serum antibody detection reagents are cheap,easy to operate and time-sav-ing,and have been widely used in China. However,these agents are not stable in sensitivity and specificity, and because of that their accuracy in the diagnosis of smear-and culture-negative TB is doubtful. In this re-view,we summarize some problems in the use of serum antibody detection among smear- and culture-nega-tive pulmonary TB patients and discuss possible methods to solve these problems expecting to provide some ideas for promoting its development,application and policy formulation.

Surveillance and analysis of virus antibodies in conventional rabbits and guinea pigs in Guangdong Province during 2014-2016 / 中国比较医学杂志

Objective To analyze the possibilities of screening SPF rabbits and guinea pigs from conventional animals, viral antibodies of the conventional rabbits and guinea pigs bred by licensed companies in Guangdong province during 2014-2016 were determined according to the standard of SPF animals in GB14922. 2. Methods Nine batches of 167 rabbit sera and 155 guinea pig sera were sampled from 6 companies. Serum antibodies to virus were determined by ELISA according to GB14922. 2. Results Positivity of antibody to rabbit hemorrhagic disease virus (RHDV) was 82. 2%(129/157) for the vaccinated rabbits, and negative result were obtained for unvaccinated rabbits. Positive rate of rabbit rotavirus (RRV) was 42. 5% (71/167). No positive antibody responses to Sendai virus were detected out in all rabbits. The positive rates of guinea pig reovirus type III (REO-3) and pneumonia virus of mice (PVM) were 52. 9%(82/155)and 20% (31/155) respectively. Antibody responses to Sendai virus ( SV) and lymphocytic choriomeningitis virus ( LCMV) were negative in all guinea pigs. Conclusions Although the conventional rabbits and guinea pigs could meet the national standard, higher infection rates of virus excluded that SPF animals emerged in conventional animals, indicating that selection of SPF animals from conventional colonies is impracticable.

Nine pathogens IgM antibody detection of 5918 respiratory tract infections in Shenzhen city / 国际检验医学杂志

Objective To analyze the infection characteristics and epidemic trend of 9 respiratory pathogens in Shenzhen popula-tion .Methods The 5918 patients with acute respiratory tract infection were collected ,indirect immunofluorescence was used to de-tect the IgM antibody of 9 respiratory pathogens ,including legionella pneumophila type 1 (LP1) ,mycoplasma pneumoniae (MP) , rickettsia Q (COX) ,rickettsia and chlamydia pneumoniae (CPn) ,adenovirus (ADV) ,respiratory syncytial virus (RSV) ,influenza A virus (INFA) ,influenza B virus (INFB) ,parainfluenza virus (PIVs) ,the results were analyzed statistically .Results A total of 1376 samples were detected at least one pathogen in 5918 serum samples ,the total positive rate was 23 .25% .The positive rate of MP was the highest (15 .19% ) ,followed by the INFB (8 .11% ) .The positive rates of other pathogens were relatively low .The mixed infection positive rate was 4 .29% .The positive rates of MP ,INFB and PIVs in women were significantly higher than those in men(P<0 .001) .The positive rates of MP ,INFB and PIVs were different in different seasons ,the differences were statistically sig-nificant(P<0 .001) .The positive rates of MP ,ADV and INFB in 0 to 14 years old group were significantly higher than those in >14 to 60 years old group and >60 years old group(P<0 .05) .The positive rates of MP ,CPn ,ADV ,RSV ,INFB and PIVs were closely related with age in infant and children (0 to 14 years old)(P<0 .05) .Conclusion The main respiratory pathogens of SARS in Shenzhen city were MP and INFB ,the 9 pathogens had their own infection characteristics and epidemic trend .

Surveillance and analysis of virus antibodies in conventional rabbits and guinea pigs in Guangdong Province during 2014-2016 / 中国比较医学杂志

Objective To analyze the possibilities of screening SPF rabbits and guinea pigs from conventional animals, viral antibodies of the conventional rabbits and guinea pigs bred by licensed companies in Guangdong province during 2014-2016 were determined according to the standard of SPF animals in GB14922. 2. Methods Nine batches of 167 rabbit sera and 155 guinea pig sera were sampled from 6 companies. Serum antibodies to virus were determined by ELISA according to GB14922. 2. Results Positivity of antibody to rabbit hemorrhagic disease virus (RHDV) was 82. 2%(129/157) for the vaccinated rabbits, and negative result were obtained for unvaccinated rabbits. Positive rate of rabbit rotavirus (RRV) was 42. 5% (71/167). No positive antibody responses to Sendai virus were detected out in all rabbits. The positive rates of guinea pig reovirus type III (REO-3) and pneumonia virus of mice (PVM) were 52. 9%(82/155)and 20% (31/155) respectively. Antibody responses to Sendai virus ( SV) and lymphocytic choriomeningitis virus ( LCMV) were negative in all guinea pigs. Conclusions Although the conventional rabbits and guinea pigs could meet the national standard, higher infection rates of virus excluded that SPF animals emerged in conventional animals, indicating that selection of SPF animals from conventional colonies is impracticable.

Nine pathogens IgM antibody detection of 5918 respiratory tract infections in Shenzhen city / 国际检验医学杂志

Objective To analyze the infection characteristics and epidemic trend of 9 respiratory pathogens in Shenzhen popula-tion .Methods The 5918 patients with acute respiratory tract infection were collected ,indirect immunofluorescence was used to de-tect the IgM antibody of 9 respiratory pathogens ,including legionella pneumophila type 1 (LP1) ,mycoplasma pneumoniae (MP) , rickettsia Q (COX) ,rickettsia and chlamydia pneumoniae (CPn) ,adenovirus (ADV) ,respiratory syncytial virus (RSV) ,influenza A virus (INFA) ,influenza B virus (INFB) ,parainfluenza virus (PIVs) ,the results were analyzed statistically .Results A total of 1376 samples were detected at least one pathogen in 5918 serum samples ,the total positive rate was 23 .25% .The positive rate of MP was the highest (15 .19% ) ,followed by the INFB (8 .11% ) .The positive rates of other pathogens were relatively low .The mixed infection positive rate was 4 .29% .The positive rates of MP ,INFB and PIVs in women were significantly higher than those in men(P<0 .001) .The positive rates of MP ,INFB and PIVs were different in different seasons ,the differences were statistically sig-nificant(P<0 .001) .The positive rates of MP ,ADV and INFB in 0 to 14 years old group were significantly higher than those in >14 to 60 years old group and >60 years old group(P<0 .05) .The positive rates of MP ,CPn ,ADV ,RSV ,INFB and PIVs were closely related with age in infant and children (0 to 14 years old)(P<0 .05) .Conclusion The main respiratory pathogens of SARS in Shenzhen city were MP and INFB ,the 9 pathogens had their own infection characteristics and epidemic trend .

Development and evaluation of an immunochromatographic assay using a gp51 monoclonal antibody for the detection of antibodies against the bovine leukemia virus

Infection of cattle with bovine leukemia virus (BLV) has been observed and reported worldwide, including in Korea. The onsite identification of infected cattle would help decreasing and eradicating BLV infections on farms. Here, we present a new immunochromatographic assay that employs monoclonal antibodies (MAbs) for the detection of antibodies against BLV in the field. BLV envelope glycoprotein (gp)51 was expressed in E. coli, and MAbs against recombinant BLV gp51 were generated for the development of an immunochromatographic assay to detect BLV antibodies in cattle. The sensitivity and specificity of the assay were determined by comparing these results with those obtained from a standard enzyme linked immunosorbent assay (ELISA). A total of 160 bovine sera were used to evaluate the new immunochromatographic assay. Using ELISA as a reference standard, the relative specificity and sensitivity of this assay were determined to be 94.7% and 98%, respectively. Because of its high sensitivity and specificity, this BLV antibody detection assay would be suitable for the onsite identification of BLV infection in the field.

Analysis of 17 175 cases with results of physical examination crowd EB virus antibody positive / 中国基层医药

Objective To study the relationship between nasopharyngeal carcinoma and positive IgA antibodies of EB virus nuclear antigen 1 (NA),EB virus capsular antigen (VCA) and EB virus Zta protein.Methods The serum EB virus VCA-IgA,NA1-IgA and Zta-IgA antibody in 17 175 cases were detected by ELISA.782 cases of EB virus antibody-positive subjects were further given nasopharyngeal CT imaging,electronic nasopharyngoscopy.Finally confirmed by biopsy and immunohistochemistry.Comparison of serology results EB antibody-positive individual,while the two antibody-positive and antibody while three positive nasopharyngeal carcinoma detection rate,combined with evaluation of EB virus antibody detection in nasopharyngeal carcinoma screening value of high-risk groups.Results 17 175 cases of medical groups,the EB virus antibodies were detected in 782 cases,with the most positive individual,accounting for 535 cases (68.41％),nasopharyngeal cancer diagnosed in 15 cases (2.80％);two antibody positive while 213 cases (27.24 ％),diagnosed 49 cases of nasopharyngeal carcinoma (23.00％);least three antibody positive while only 34 cases (4.35％),nasopharyngeal cancer diagnosed 18 patients (52.94％);by physical examination,in 16,393 cases of EB virus antibody negative population by nasopharyngoscopy and throat CT and other medical examination,the final diagnosis of nasopharyngeal carcinoma in 6 cases (0.04％).Conclusion Three EB virus antibody combined detection greatly improves the rate of nasopharyngeal cancer diagnosis;we must also do other physical examination to prevent EB virus antibody-negative nasopharyngeal crowd missed.

Analysis of HIV antibody screening results from 2013 to 2015 in Qinnan District of Qinzhou / 国际检验医学杂志

Objective To analyze the data of Human Immunodeficiency Virus (HIV ) screening in Qinnan District of Qinzhou from 2013 to 2015 ,and to provide scientific evidences for the prevention and control of Acquired Immune Deficiency Syndrome (AIDS) .Methods HIV antibody data which was collected from the AIDS laboratory between 2013 and 2015 in Qinnan District of Qinzhou were analyzed .Results Of 3 955 cases of the HIV antibody detecting in the AIDS laboratory from 2013 to 2015 ,200 cases were HIV antibody positive ,and the positive rate was 5 .06% which was decreased compared with previously .The HIV antibody positive rate of man was 9 .26% ,which was higher than women .Among the HIV positive people ,the rate of the ages older than 50 was 25 .36% which was the highest compared to the other age groups .The positive rate of farmers was 30 .83% and the rate of di‐vorced or widowed was 30 .86% and the rate of the illiterate was 16 .08% .Conclusion Man ,farmers ,the age older than 50 years old ,the divorced or widowed and the low degree of education groups are the focus groups of health education and prevention of AIDS .If we can find HIV infectors earlier ,we can control the epidemic of AIDS and formulate the prevention and control strategies better .

Cloning and expression of MICB gene and its application for the detection of anti-MICB antibodies / 中华微生物学和免疫学杂志

Objective To construct a prokaryotic expression system for major histocompatibility complex class Ⅰ chain-related gene B ( MICB) and to establish an ELISA method for the detection of anti-MICB antibodies in patients with kidney transplantation. Methods The MICB cDNA fragments were ob-tained by RT-PCR with a pair of specific primers. The MICB cDNA and the prokaryotic expression vector pET-28a were digested by two restriction enzymes and ligated to construct the recombinant expression plas-mid pET-28a-MICB. The transformed E. coli BL21 DE3 strains carrying recombinant expression plasmid were induced by IPTG to express MICB protein. The expressed recombinant proteins were identified by Western blot assay and purified by Ni-NTA Spin column. The purified proteins were coupled to ELISA for the detection of anti-MICB antibodies in patients with kidney transplantation. Results Three common MICB fragments contained the exons 2 and 3 were obtained. The recombinant proteins were expressed in E. coli BL21 DE3 strains carrying pET-28a-MICB and successfully purified by the Ni-NTA Spin column. Results of the Western blot assay confirmed that the obtained proteins were the target proteins. The ELISA method was successfully established and used for the detection of anti-MICB antibodies in 24 patients with kidney trans-plantation. The absorbance values indicated that the sensitivities of three recombinant MICB proteins were different. Conclusion The expression system for MICB gene was successfully constructed. The established ELISA for the detection of anti-MICB antibodies would pave the way for further investigation on the correla-tions between MICB protein and transplantation immunity.

Application of nucleic acid detection and antibody detection in HIV infection / 国际检验医学杂志

Objective To compare the accuracy of nucleic acid detection and antibody detection for diagnosing human immunode‐ficiency virus(HIV) infection .Methods Retrospectively analysed data of nucleic acid detection and antibody detection from 124 ca‐ses of patients diagnosed with HIV infection from 2005 to 2014 .The positive rates of the two methods were compared respectively in patients with early‐stage of HIV infection(76 cases) and patients with intermediate and advanced stage of HIV infection (48 ca‐ses) .Results In patients with early‐stage of HIV infectionn ,the positive rate of nucleic acid detection (94 .74% ) was higher than that of antibody detection (84 .21% );while in patients with intermediate and advanced stage of HIV infection ,the positive rate of antibody detection(97 .92% ) was higher than that of nucleic acid detection (81 .25% );both had statistically significant difference (P<0 .05) .Conclusion On the early stage of HIV infection ,the accuracy of nucleic acid detection is higher than that of antibody detection ;while on the intermediate and advanced stage of HIV infection ,antibody detection shows better accuracy .