1.Applicability evaluation of different ELISA kits for SARS-CoV-2 antibody screening of post-immunization plasma
Chinese Journal of Biologicals 2023;36(12):1476-1482+1490
SARS-CoV-2;ELISA-IgG;Neutralizing antibody;Dilution multiple;Post-immunization plasma;Qualified rate of plasma collection
Objective To determine the optimal plasma screening kit for raw materials by comparing the qualified rate of plasma collection and operation convenience of four SARS-CoV-2 ELISA-IgG and neutralizing antibody kits from three manufacturers.MethodsSingle plasma samples with different ELISA dilution multiples screened from existing plasma immunized with SARS-CoV-2 vaccine were used as raw materials,which were detected by using SARS-CoV-2 ELISA-IgG and neutralizing antibody kits from different manufacturers. Plasma samples were mixed according to the potency level of each manufacturer,and pseudovirus neutralization test was carried out on pooled plasma. According to the potency of neutralizing antibody,the plasma collection standard of different kits were determined,and the qualified rate of plasma collection was calculated respectively. The dilution method of single plasma sample screening was determined in regard of the plasma collection standard,and the operation convenience was compared;The applicability of the kits were evaluated comprehensively.ResultsThe qualified rate of ELISA-IgG antibody kit of manufacturer B in 1 108 WT > 16 plasma samples was 82. 1% with internal control reference B2 as the plasma collection standard,which was higher than that of the current ELISA-IgG antibody kit of manufacturer A(59. 8%). With the level of 50 IU/mL tested by neutralizing antibody kit of manufacturer B as the standard of plasma collection,the qualified rate of plasma collection of this kit in 387 WT16 ~64 samples was 66. 4%,higher than that of ELISA-IgG kit of manufacturer A(42. 1%). While with the level of 100 IU/mL tested by neutralizing antibody kit of manufacturer C as the standard of plasma collection,the qualified rate of plasma collection of this kit in 536 WT16 ~ 64 samples was 42. 5%,lower than that of ELISA-IgG kit of manufacturer A(50. 2%).Neutralizing antibody kit of manufacturer C had the most operation steps and the longest reaction time;ELISA-IgG kit of manufacturer B had the shortest reaction time,and required no out-of-hole dilution for samples of WT16 ~ 64,while the ELISA-IgG kit of manufacturer A required out-of-hole dilution in transition plate. The difference of qualified rate of plasma collection among the four kits mainly occurred in the samples of WT16 ~ 64. The order of the qualified rate of plasma collection in this range was:ELISA-IgG kit of manufacturer B > neutralizing antibody kit of manufacturer B > ELISA-IgG kit of manufacturer A > neutralizing antibody kit of manufacturer C. The order of operation convenience was:ELISA-IgG kit of manufacturer B > neutralizing antibody kit of manufacturer B > ELISA-IgG kit of manufacturer A > neutralizing antibody kit of manufacturer C.ConclusionIn view of the overall analysis of the applicability from qualified rate of plasma collection and operation convenience,ELISA-IgG kit of manufacturer B is the optimal.
2.Optimization,verification and application of ELISA method for quantitative detection of varicella-zoster virus IgG antibody
Chinese Journal of Biologicals 2023;36(8):973-979
Objective To optimize and verify the ELISA method for quantitative detection of varicella-zoster virus(VZV)IgG antibody potency,and use it for the screening of plasma with high potency VZV-IgG in healthy donors.Methods The VZV-IgG indirect ELISA kit from Institut VirionSerion GmbH was selected,the first international standard for varicellazoster immunoglobulin(NIBSC code:W1044)was diluted to 2 IU/mL as the standard,and 4-parameter fitting curve was used to develop the quantitative ELISA method. The method was determined for the optimal linear range and verified for the precision and accuracy. VZV-IgG antibody potency of 1 962 human plasma samples and some batches of human immunoglobulin preparations from 10 plasma stations under Sinopharm Wuhan Plasma-derived Biotherapies Co.,Ltd.(SWPB)were detected by the developed method.Results The linear range of the standard curve was 16. 25 ~ 2 000 mIU/mL,the CV values of precision in intra-and inter-assays were 1. 3% ~ 10. 6% and 4. 270% ~ 7. 636%,and the accuracy in intra-and inter-assays were 92. 30% ~ 111. 02% and 98. 40% ~ 104. 88%,respectively;Sample-adding experiment showed that the measured value of the added sample was 95. 79% ~ 111. 03% of the theoretical value. The positive rate of 1 962 human plasma samples was 94. 29%,and the samples with potency greater than 3 000 mIU/mL accounted for 1. 02%. The potency of VZV-IgG antibody in different kinds of human immunoglobulin preparations was lower,while higher than that of intravenous human immunoglobulin(pH 4).Conclusion The optimized VZV-IgG quantitative detection method can be used for the screening of VZV-IgG in healthy people. The positive rate of VZV-IgG antibody in naturally infected healthy plasma donors is high,while the potency is low,thus,vaccine immunization is required to obtain qualified plasma with high potency.
3.Preparation of the second generation internal control reference for anti-SARS-CoV-2 IgG antibody and evaluation of its applicability in ELISA detection method
Chinese Journal of Biologicals 2023;36(9):1085-1092
Objective To prepare the second generation internal control reference(B2)for Ig G antibody against severe acute respiratory symptom coronavirus 2(SARS-CoV-2)and evaluate its applicability in ELISA detection method. Methods Among the volunteers vaccinated with SARS-CoV-2 inactivated vaccine(BBIBP-Cor V)produced by Beijing Institute of Biological Products Co.,Ltd.,19 Ig G antibody positive plasma samples with ELISA-Ig G dilution ratio of 20 ~ 60 were screened,and the Ig G antibody,IgM antibody and neutralizing antibody were detected by ELISA,B2 was prepared from nonlipid plasma with ELISA-Ig G dilution ratio of 32 ~ 45,IgM negative and similar neutralizing antibody inhibition rate. The neutralizing antibody potency of the first generation internal control reference(B1)and B2 detected by ELISA was calibrated with the first generation WHO international standard of anti-SARS-CoV-2 immunoglobulin(NIBSC 20/136),and the accelerated stability(storage at 2 ~ 8 ℃ for 5,8,14,20,and 30 d respectively),the service stability(storage at 18 ~25 ℃ for 1,2,and 3 h respectively),the freeze-thaw stability(1,2 and 3 times)and the long-term stability(storage at-25 ℃ for10 months)of B2 were tested. B2 was used as standard to detect plasma after single vaccine immunization and mixed plasma was prepared according to different ELISA-Ig G dilution ratio. The correlation and linear regression analysis between ELISA-Ig G dilution ratio and neutralizing antibody potency of pseudovirus in mixed plasma were carried out. Results Among 19 plasma samples,5 samples were non-lipid plasma with ELISA-Ig G dilution ratio of 32 ~ 45,IgM negative and similar neutralizing antibody inhibition rate. B2 was prepared by mixing every plasma in equal volume fraction,and the dilution ratio of ELISA-Ig G was assigned to 32. The neutralizing antibody potency of B1 calibrated with NIBSC 20/136 was 133. 38 EIU/m L and that of B2 was 122. 14 EIU/m L. The recovery rates of accelerated stability,service stability,freeze-thaw stability and long-term stability of B2 were all in the range of(100 ± 15)%. The ELISA-Ig G dilution ratio of the mixed plasma from the same source was significantly correlated with the neutralizing antibody potency of pseudovirus.(each R~2> 0. 99,each P < 0. 000 1).Conclusion B2 prepared from plasma immunized with SARS-CoV-2 inactivated vaccine can replace B1 prepared from plasma of COVID-19 convalescent patients.
4.Clinical observation on Yiyi Fuzi Baijiang powder for treatment of patients with ulcerative colitis accompanied by yang-deficiency of spleen and kidney: a report of 80 cases
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2017;24(4):419-422
Objective To observe the clinical effects of Yiyi Fuzi Baijiang powder for treatment of patients with ulcerative colitis (UC) accompanied by yang-deficiency of spleen and kidney.Methods Eighty patients with UC admitted to the First Affiliated Hospital of Tianjin University of Traditional Chinese Medicine (TCM) from January 2012 to December 2015 were enrolled, and they were divided into an integrated TCM and western medicine group (combined group) and a control group by random number table method, 40 cases in each group. The control group was treated by normal saline 100 mL with dexamethasone 10 mg enema, whilethe combined group was given Yiyi Fuzi Baijiang powder 100 mL (coix seed 30 g, aconitum carmichaeli debx 6 g, herba patriniae 15 g, radix paeoniae alba 15 g, frankincense 6 g, myrrh 6 g, glycyrrhiza uralensis 6 g) combined with dexamethasone 10 mg enema, one therapeutic course was 10 days, and after 3 courses, the clinical efficacy was observed in the two groups. The differences of erythrocyte sedimentation rate (ESR), the number of red blood cell (RBC) and white blood cell (WBC) positive patients in stool routine were compared before and after treatment, and the clinical therapeutic effects were observed in the two groups.Results After treatment, the ESR was reduced in both groups compared to that before treatment, the numbers of RBC and WBC positive patients in stool routine were decreased in both group compared to those before treatment, and the degrees of changes were more significant in combined group than those in the control group [ESR (mm/1 h): 15.00±3.07 vs. 20.55±3.87, the number of RBC positive patients: 7 cases vs. 21 cases, the number of WBC positive patients: 14 cases vs. 21 cases, allP < 0.05]. The total effective rate of combined group was significantly higher than that of the control group [92.5% (37/40) vs. 75.0% (30/40),P < 0.05].Conclusion The clinical effect of Yiyi Fuzi Baijiang powder combined with dexamethasone enema was superior to that simply using hormone enema therapy for treatment of patients with UC accompanied by yang deficiency of spleen and kidney, this combined method is simple, and its curative effect is definite.
5.An observation on clinical efficacy of paeoniae decoction add and subtract combined with mesalazine for treatment of ulcerative colitis
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2017;24(3):239-242
Objective To investigate the clinical efficacy of paeoniae decoction (PD) add and subtract or modified PD combined with mesalazine for treatment of patients with ulcerative colitis (UC).Methods A prospective study was conducted, 98 patients with UC admitted to the First Affiliated Hospital of Tianjin University of Traditional Chinese Medicine from August 2015 to August 2016 were enrolled, and they were divided into an observation group and a control group by random number table, 49 cases in each group. The control group was treated with mesalazine 1 g, 3 times daily; the observation group was treated with paeoniae decoction (ingredients: paeonia 30 g, coptis chinensis 15 g, Chinese angelica 15 g, betel nut 6 g, Chinese rhubarb 6 g, baikal skullcap 6 g, cinnamon 6 g, licorice 6 g) add and subtract, on the basis of treatment of the control group ,1 dose daily, each dose was decocted 2 times and warm decoction was taken once in the morning and once in the evening; the therapeutic course in both group was consecutive 6 weeks. The contents of different inflammatory factors, interleukin (IL-1, IL-8), tumor necrosis factor-α (TNF-α), C-reactive protein (CRP)in serum were compared before and after treatment between the two groups, and the clinical curative effects and adverse reactions were observed in two groups.Results After treatment, the levels of serum IL-1, IL-8, TNF-α, CRP in two groups were significantly lower than those before treatment, and the degrees of decrease were more significant in the observation group than those in the control group [IL-1 (ng/L): 8.48±3.05 vs. 9.38±3.37, IL-8 (ng/L): 10.15±2.23 vs. 11.94±2.30, TNF-α (μg/L): 122.13±6.40 vs. 137.02±7.35, CRP (mg/L): 7.16±1.93 vs. 8.02±2.63, allP < 0.05]. After treatment, the total effective rate in observation group was significantly higher than that in control group [91.84% (45/49) vs. 77.55% (38/49),P < 0.05]; the adverse reactions of observation group and control group had no statistical significant difference [4.08% (2/49) vs. 6.12% (3/49),P > 0.05].Conclusion The clinical efficacy of add and subtract of paeoniae decoction combined with mesalazine for treatment of ulcerative colitis is prominent, and no increase of adverse reaction occurrence was seen.
6.Searching of specialized social practice in full-time MPH program
Chinese Journal of Medical Education Research 2013;(7):660-662,663
The article reviewed of the origin of the social practice teaching in master of public health(MPH) program and gave the introduction of the social practice in American MPH education. Through the definition, positioning and feedback of specialized social practice in full-time MPH program, we can get some experiences and enlightenments of specialized social practice in full-time MPH educa-tion, which will provide references for social practice teaching in full-time professional degree education.
7.Effects of optional cesarean section on the level of plasma motilin and serum gastrin in neonatal umbilical blood
Zhijun XIA ; Shulan ZHANG ; Zhuo ZHOU
Chinese Journal of Practical Gynecology and Obstetrics 2001;17(2):107-108
ObjectiveTo explore the effects of optional cesarean section on the level of plasma motilin and serum gastrin in umbilical blood and their relationship with gastroenteric symptoms of neonates. MethodsWe measured the concentrations of plasma motilin and serum gastrin in umbilical blood from neonates undergoing optional cesarean operation, emergency cesarean section and vaginal delivery by radioimmunoassay(RIA). ResultsFull-term deliveries undergoing optional cesarean section were with significant gastrointestinal symptoms. Plasma motilin and serum gastrin were significantly lower in neonates undergoing cesarean operation without birth initiate group(optional cesarean section)than that in cesarean section with birth initiate group and vaginal delivery group (P<0.05). ConclusionCesarean operation without birth initiate influences neonatal gastroenteric symptoms.Breast feeding which begins as early as possible will help to relieve those symptoms.
9.Advances in treatment of retinopathy of prematurity
Yu WANG ; Xiaoguang ZHOU ; Zhijun CHEN
Chinese Journal of Applied Clinical Pediatrics 2017;32(2):146-148
Retinopathy of prematurity (ROP) is a potentially blinding retinal vascular disease that occurs in premature infants and low birth weight infants.Along with the development of perinatal medicine,the incidence of ROP has increased.If infants with ROP could get treatment in time,the disability rate due to ROP would significantly reduce.Now,the present situation and advances in treatment of ROP were reviewed.
10.Formulation of local standard of Microtus Fortis as a laboratory animal:formulation and related research
Zhijun ZHOU ; Zhijie SU ; Yuanjing YU
Acta Laboratorium Animalis Scientia Sinica 2014;(6):103-105
The principle, basis, necessity and significance of formulating the local standard of Microtu fortis as a laboratory animal were described in this paper, and the standard was compared with the relationship between this standard of Microtu fortis as laboratory animal and the existing laws, regulations of other standards of laboratory animals.The specific procedures and the degree of adoption of domestic standards and advanced foreign standards were introduced.Furthermore, the proposal and the reasons of recommendatory standards were presented.