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【Objective】 To analyze the results of different methods for reactive samples screened by the enzyme linked immunosorbent assay (ELISA) against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in blood donors. 【Methods】 From March to April 2020, a total of 8 632 blood samples in Shenzhen were screened for SARS-CoV-2 total antibodies (TAb, including IgG, IgM, IgA) in plasma using ELISA(PC group), the antibody reactivity samples and their follow up plasma samples (FC group), and samples of disease control group(DC group) from January to April 2020 were detected using the following methods: 1) ELISA method for detecting IgG, IgM, and (or without detection) TAb; 2) pseudovirus neutralizing antibody test(pVNT); 3) western blot (WB) of SARS-CoV-2 antibody. The negative control group(NC group) from February to April 2020 performed ELISA and WB testing. 【Results】 Among the 34 total antibody positive samples, 2 were positive for pVNT test, and the total antibody, IgG and WB in the initial screening and tracking testing were positive. Thereafter, it was determined to be confirmed positive. The other 2 cases were positive for pVNT test, while the samples with positive WB results were in the follow-up stage. The TAb, IgG, and pVNT results did not conform to the dynamic evolution of antibodies, and cannot be determined as confirmed positive. 【Conclusion】 The infection status of antibody reactivity samples screened by SARS-CoV-2 ELISA can be judged by the logic of pVNT, WB and the dynamic change of antibody.
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【Objective】 To investigate the confirmatory status of HIV-1 antibody detection and Western blot (WB) test among voluntary blood donors in Wuhu, and to explore the strategies and methods to further ensure blood quality and safety. 【Methods】 Blood samples were preliminarily screened by ELISA and NAT, and the reactive samples were sent to Wuhu CDC for further WB test of HIV-1 antibody. The confirmation results of HIV-1 antibodies of voluntary blood donors in Wuhu in the past 10 years were retrospectively collected. The characteristics of WB bands of positive samples were analyzed, and the demographic characteristics of HIV-infected voluntary blood donors were sorted out. 【Results】 A total of 354 864 blood samples from voluntary blood donors in Wuhu during January 2011 to May 2021 were investigated, among which 42 were confirmed HIV positive (HIV-1 antibody positive in 41, and solo HIV-RNA reactive in 1), with a total HIV positive rate of 11.8/100 000(42/354 864). Statistical differences were found in gender [males 97.6% (41/42) vs females 2.4% (1/42)], marital status [unmarried 17.3/100 000 vs married 8.0/100 000] and occupation [staff/workers 37.5/100 000 vs students11.4/100 000 vs others 7.7/100 000]. Among the positive samples, the yield rate of WB bands gp160 was 100% (41/41), both gp41 and p24 were 97.6% (40/41),, and p55 was the lowest 46.3% (19/41). P51 and P66 presented the highest yield consistency (Kappa=1.000, P5 000 cps/mL by viral load (VL) testing, indicating HIV window period infection. 【Conclusion】 HIV infection statistically affected male donors more than females in Wuhu area, and most were early infection that revealed by WB band analysis. NAT plays an important role in the detection and confirmation of HIV infection during the window period, and is essential for blood safety.
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Objective To analyze the positive results of HIV antibody screening in the laboratory of AIDS confirmation center of Hubei Provincial Center for Disease Control and Prevention from 2014 to 2020, and to provide a basis for improving detection strategies. Methods A total of 2 728 primary screening positive specimens received by the laboratory of Hubei confirmation center from 2014 to 2020 were retested with two reagents. Specimens with at least one reactive result were confirmed with western blot (WB). The samples with uncertain or negative WB results were further confirmed by nucleic acid quantitative detection. The test results were analyzed retrospectively. Results A total of 2 297 specimens with positive retest results were confirmed by WB, with a positive rate of 93.47%. The highest proportion of patients was from medical institutions. The positive rate detected by 4 diagnostic kits was apparently higher in S/CO＞10 cases than that in S/CO≤10, and the difference was statistically significant (P 5 000cps / ml, and 12 cases were TND. 13 of the 30 WB negative samples had nucleic acid test results＞5 000CPs/mL . Conclusions The coverage of HIV screening laboratories in hospitals at all levels should be further increased to find more HIV infected persons. The anti-HIV ELISA S/CO ratio is correlated with the positive results confirmed by western blot. Therefore, ELISA S/CO ratio can be used to predict anti-HIV antibody positivity. For samples with uncertain or negative WB detection, supplemental nucleic acid test should be carried out timely for early diagnosis.
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Objective To analyze the results of HIV antibody screening test and confirmation test results in Pudong New Area from 2014 to 2018, and to understand the situation of HIV infection in the region in recent years. Methods Samples that responded to initial screening of HIV in this jurisdiction area were tested by enzyme-linked immunosorbent assay (ELISA), chemiluminescence assay or immunochromatographic rapid diagnosis test. The samples that showed a positive response to one of the reexamination methods were subjected to confirmatory tests by Western blot assay. EXCEL and SPSS 16.0 statistical software were used to collate and analyze the data. Results Confirmatory tests were performed on a total of 2 100 retested samples. HIV positive samples were 1485 (70.7%), and HIV negative and uncertainty samples were 452 (21.5%) and 163 (7.8%), respectively. The confirmed positive male/female sex ratio was 7.95:1, and the difference was statistically significant (χ2=3.745, P<0.001). The age of confirmed positive patients was mainly 15~39 years old. The proportion of voluntary testing and counseling was the highest among the population receiving HIV tests. The positive coincidence rate between the review test and confirmation test was 91.3%. The gp160 and gp120 bands appeared most frequently in WB-confirmed positive samples, which was 100%. The gp160 band was the most frequent band of uncertain samples, accounting for 71.4%. Conclusion The positive rate was high in the confirmatory test of samples with positive results by both methods, and the samples with one negative and one positive were mostly uncertain or negative in the confirmatory test. When the results were uncertain, CD4, nucleic acid detection, and viral load detection should be used as auxiliary diagnosis.
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Objective To investigate the band pattern characteristics of Western bolt (WB) by analyzingthe change of anti-virus antibodies from patients with different clinical stages of human immunodeficiency virus 1 (HIV-1) infection.Methods Antibodies were detected by WB test,CD4+ T cells by flow cytometer.Results Among 208 cases,193 male cases accounted for 92.79％,and 163 cases of men who have sex with men (MSM) accounted for 78.37％.The antibodies against viral protein gp160,gp120,gp41,p66,p51,p31,p24 showed high positive rates,and no significant different was detected across different clinical stages.The positive rate of p55 antibody,encoding by gag gene,was significantly higher in those HIV positive individuals infected via homosexual compared with HIV-1 positive individuals through heterosexual contact (P＜0.05).The 5 common WB bands patterns were as following:all bands①,missing p55②,missing p39③,missing p55 + p39④ and missing p39 + p55 + p17⑤.Primary stage infection group showed the highest appearance rate of all bands (44.2％).Conclusions The WB bands patterns and CD4+ T lymphocyte counts can help to judge immune status,determine disease stages and monitor disease progression.We should take effective measures for the MSM and floating population.
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Objective To compare the differences of two kinds of EIA reagents for HIV-1/2 (Ab/Ag) screening results of voluntary blood donors,in addition to find out the feasibility of reducing 1 times of EIA detection.Methods To collect data of HIV 1/2 screening positive results and confirmatory test for voluntary blood donors from 2009 to 2014 in Jiaxing area,and to compare the relationship of screeing test results with that of the confirmatory test,and then to analyze the relevance between S/CO values of screening test and confirmatory test.Results Screening positive rates of domestic and imported reagents,which were 9.58/10 000 and 12.43/10 000,respectively;and the confirmatory coincidence rates were 11.84％ and 9.12％,respectively.There was no significant difference (x2 =1.11,P＞0.05).The double-reagent joint detection positive rate was 1.37/10 000,and its positive predictive value was 82.86％.Single-reagent test result compared with that of double-reagent test,which had significant differences (x2domestic =94.04,P＜0.05 and x2ximported =124.86,P＜0.05).When the S/CO value was more than 6,domestic and imported reagents positive predictive values were 93.55％ (29/31) and 87.50％ (28/32),respectively.Conclusion There is no difference between domestic and imported reagents EIA-HIV1/2.
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Objective To investigate the band pattern characteristics of Western bolt (WB) by analyzingthe change of anti-virus antibodies from patients with different clinical stages of human immunodeficiency virus 1 (HIV-1) infection.Methods Antibodies were detected by WB test,CD4+ T cells by flow cytometer.Results Among 208 cases,193 male cases accounted for 92.79％,and 163 cases of men who have sex with men (MSM) accounted for 78.37％.The antibodies against viral protein gp160,gp120,gp41,p66,p51,p31,p24 showed high positive rates,and no significant different was detected across different clinical stages.The positive rate of p55 antibody,encoding by gag gene,was significantly higher in those HIV positive individuals infected via homosexual compared with HIV-1 positive individuals through heterosexual contact (P＜0.05).The 5 common WB bands patterns were as following:all bands①,missing p55②,missing p39③,missing p55 + p39④ and missing p39 + p55 + p17⑤.Primary stage infection group showed the highest appearance rate of all bands (44.2％).Conclusions The WB bands patterns and CD4+ T lymphocyte counts can help to judge immune status,determine disease stages and monitor disease progression.We should take effective measures for the MSM and floating population.
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Objective:Establishing the hepatic stem cell(WB-F344)line that expresses human interleukin-2 stably and effectively.Methods: We constructed the retroviral vector Plpcx-il-2 containing human interleukin-2 gene, and transfected it into PT67 packaging cells.The infectious viruses were used to infect the hepatic stem cell WB-F344,then the puromycin resistant clones were acquired after puromycin selection.The expression of IL-2 was detected using RT-PCR,immunocytochemistry, Western blot and indirect immunofluorescence, respectively.Results: The retroviral vector containing human IL-2 gene was constructed.The titer of virus in the supernatant reached 10~5 CFU/ml after transfecting PT67 packeting cells.The puromycin resistant clones were acquired after infection of WB-F344 and the cell line WB-F344/Plpcx-IL-2 expressing human interleukin-2 stably was established.Conclusion: By retroviral-mediated transfecting human interleukin-2 gene, the hepatic stem cell stably and effectively expressing interleukin-2 are obtained.This provides a good basis for further research on immuno-gene therapy of liver cancer.
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Objective: To investigate the effect of sodium butytate (different concentrations) on the growth and proliferation of rat liver oval cell line WBF344, and to discuss the conditions and tules for sodium butyrate-inducd WB-F344 cells differentiation into biliary lineage in vitro. Methods: WB-F344 cells were treated with sodium butyrate (0.70, 2.25, 3.75, 4.5 mmol/L) and the cell growth and morphological changes were observed; routinely cultured WB-F344 cells were taken as control. The changes of CK19 protein expression were examined immunohistochemically after WB-F244 cells were treated with 3.75% sodium butyrate; and the expression of phenotypic markers, such as γ-glutamyltransferase (GGT), β4-integrin, CK19, AFP and ALB at toRNA level were determined by RT-PCR. Untreated WB-F344 cells were used as blank control. Results: We found that sodium butyrate inhibited the growth of WB-F344 cells. The optical densities were significantly decreased in 3.75 and 4.5 mmol/L groups compared with that in control group(P<0.01); but no significant difference was found between 0.75, 2.25 mmol/L groups with control group. WB-F344 cells treated with 3.75, 4.5 mmol/L sodium butyrate became larger and round, with increased nuclei and decreased nucleus to cytoplasm ratio; those treated with 0.75, 2.25 mmol/L had no obvious changes. Immunohistochemical results showed that sodium butyrate significantly increased CK19 expression compared with control group ([92.3±1.1]% vs [1.3±0.2]%, P<0.01). RT-PCR showed increased expression of β4-integrin in sodium butyrate treated groups, but not in control group; the expression of GGT and CK19 was higher than that of control group. Alpha-fetoprotein (AFP) expression was observed in blank control group, but not in sodium butyrate treated cells. Albumin expression was not detected in the 2 groups. Conclusion: Sodium butyrate at 3.75 mmol/L is suitable for inducing WB-F344 cells differentiate into the biliary lineage in vitro.
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O Brasil é o país com o maior número de pessoas infectadas pelos vírus linfotrópicos de células T humanas dos tipos 1e -2 (HTLV-1 e HTLV2) com mais de 2,5 milhões de indivíduos infectados. Em 1993, a realização de testes sorológicos específicos tornou-se obrigatória em Bancos de Sangue. O HTLV-1 causa leucemia/linfoma de células T do adulto e mielopatia associada ao HTLV-1/paraparesia espástica tropical além de outras doenças, enquanto o HTLV-2 pode causar alguns quadros neurológicos e alterar a evolução de HIV/Aids. Os testes sorológicos que identificam anticorpos específicos disponíveis no mercado têm falhado no diagnóstico, principalmente de infecção por HTLV-2. Vários algoritmos de testes de triagem e confirmatórios têm sido propostos, mas nenhum deles se mostrou 100% eficiente com casuística de alto risco. Muitos soros resultam em padrão indeterminado no Western blot, e os isolados virais utilizados na composição dos kits podem ser a causa desses resultados. As técnicas de biologia molecular têm sido descritas como testes confirmatórios, mas não têm sido empregadas na rotina. Desde 1991, a Seção de Imunologia do Instituto Adolfo Lutz tem estudado a infecção por HTLV-1/2, contribuindo para o diagnóstico sorológico e molecular, e tem como desafio implantar um teste laboratorial capaz de detectar infecção causada por cepas brasileiras de HTLV-2.
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Clinical Laboratory Techniques , Polymerase Chain Reaction , Serology , Immunoenzyme Techniques , Human T-lymphotropic virus 1 , Blotting, WesternABSTRACT
During autopsy different viscerae are collected and preserved mainly for chemical examination to ascertain the cause of death. But many a times viscerae is preserved even in cases where cause of death is known. Police and prosecution side often question the merit of preservation of viscerae in such cases because defence lawyer often take this as opportunity to delay the prosecution till the Chemical Examiner’s report is available. Most of the time Chemical Examiner’s report is much delayed and sometimes West Bengal State Forensic Science Laboratory even refuse to accept the viscerae for chemical examination particularly where cause of death is given in P.M.report. In N.R.S. Medical College, Kolkata center around 3500 post mortem examinations are being done annually, on requisitions from Kolkata Police [K.P.], West Bengal Police [WBP] and General Railway Police (GRPS). Kolkata Police in almost all their requisitions request to preserve viscera and other medicolegal articles. An analysis was done on police requisition paper and autopsy report to find out the logical scientific solution.
Subject(s)
Autopsy , Cause of Death , Coroners and Medical Examiners/legislation & jurisprudence , Humans , India , Lawyers/legislation & jurisprudence , Organ Preservation , Viscera/pathologyABSTRACT
Objective:To investigate the effect of TGF-? on the expression of type Ⅰ inositol 1,4,5-triphosphate receptor in WB rat liver epithelial cells.Methods:The expression of type Ⅰ inositol 1,4,5-triphosphate receptor protein and mRNA were detected by Western blot and RT-PCR in WB rat liver epithelial cells in vitro stimulating with TGF-?.Results:The expression level of type Ⅰ inositol 1,4,5-triphosphate receptor protein and mRNA in WB rat liver epithelial cells were both increased after stimulating with TGF-? in several time points and reached the highest at 8 h stimulated, and then decreased.Conclusion:TGF-? enhance the IP_3R1 protein and mRNA expression in WB rat liver epithelial cells.
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With use of ~(125)I-udR release test,a significant natural killer activityagainst K_(562) target cells has been determined in human peripheral wholeblood(WB).r There was a gradual decrease in NK activity as rise inthe dilution or WB.In comparison with peripheral monouclear cells(PBMC)oneself,the NK activity lowered distinctly at both 1:4 and 1:8 dilution ofWB(p0.4).It seems that neither the red blood cells nor thePlasma ffeect on NK activity of WB,The NK activity in WB as well as inPBMO was improved by incubation with human leukocyte interferon.TheNK activity of WB and PBMC in patients with hepatocellular carcinomawas signiticantly lower than in normal controls.The results showed tahtNK activity may be determined by using properly diluted WB instead orPBMC,and the assay can be easily performed in clinical practice.
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Objective:To investigate the effect of sodium butytate (different concentrations) on the growth and proliferation of rat liver oval cell line WB-F344, and to discuss the conditions and rules for sodium butyrate-inducd WB-F344 cells differentiation into biliary lineage in vitro. Methods: WB-F344 cells were treated with sodium butyrate (0.75, 2.25, 3.75, 4.5 mmol/L) and the cell growth and morphological changes were observed; routinely cultured WB-F344 cells were taken as control. The changes of CK19 protein expression were examined immunohistochemically after WB-F244 cells were treated with 3.75% sodium butyrate; and the expression of phenotypic markers, such as ?-glutamyltransferase (GGT) ,?4-integrin, CK19, AFP and ALB at mRNA level were determined by RT-PCR. Untreated WB-F344 cells were used as blank control. Results: We found that sodium butyrate inhibited the growth of WB-F344 cells. The optical densities were significantly decreased in 3.75 and 4.5 mmol/L groups compared with that in control group(P