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Levels of soluble delta-like ligand 1 in the serum and cerebrospinal fluid of tuberculous meningitis patients

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Neural Regeneration Research 2012年第11期7卷 874-878页

作者： Jinghong Li Jinyi Li Yanjie Jia Department of Neurology First Affiliated Hospital of Zhengzhou University Zhengzhou 450052 Henan Province China

In this study, the levels of soluble delta-like ligand 1 in cerebmspinal fluid and serum of 50 patients with tuberculous meningitis, 30 patients with viral meningitis, 20 patients with purulent meningitis and 40 subjects without central nervous system disease were determined using an enzyme-linked immunosorbent assay. The mean levels of soluble delta-like ligand 1 in both cerebrospinal fluid and serum from patients with tuberculous meningitis were significantly higher compared with those from patients with viral meningitis or purulent meningitis or from subjects without central nervous system disease. Meanwhile, the level of soluble delta-like ligand 1 gradually decreased as tuberculous meningitis patients recovered. If patients deteriorated after treatment, the level of soluble delta-like ligand 1 in cerebrospinal fluid gradually increased. There was no correlation between the level of soluble delta-like ligand 1 and the protein level/cell number in cerebrospinal fluid. Our findings indicate that the levels of soluble delta-like ligand 1 in cerebrospinal fluid and serum are reliable markers for the diagnosis of tuberculous meningitis and for monitoring treatment progress. At the same time, this index is not influenced by protein levels or cell numbers in cerebrospinal fluid.

关键词： delta-like ligand 1 cerebrospinal fluid enzyme linked immunosorbent assay tuberculous meningitis

Evaluation on Clinical Application of Three Testing Methods for Human Cytomegalovirus Infection in Pregnancy

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2003年第2期23卷 192-194页

作者：曾万江闻良珍陈素华凌霞珍 Department of Obstetrics and Gynecology Tongji HospitalTongji MedicalCollegeHuazhong University of Science and Technology Wuhan 430030

The value of ELISA, N-PCR and RT-PCR in clinical practice for pregnant women with HCMV infection was investigated. 5581 pregnant women were screened by ELISA. Among them, 100 cases were positive for IgM (group 1). 69 for both IgM and serous DNA (group 2) and 69 for both IgM and mRNA (group 3). The infectious status, maternal-fetal transmission and pregnancy outcome were monitored. It was demonstrated that the accordance rate of group 3 and group 2 with group 1 was 56. 25 % and 43. 75 % , respectively. The maternal-fetal transmission rate in the group 1, 2 and 3 was 19. 00 % , 40. 58 % and 46. 15 %, respectively, with a significant difference found between group 2, 3 and group 1 (P<0. 01). Incidence of spontaneous abortion, fetal death, fetal abnormality and neonatal death in group 1, 2 and 3 was 10. 00 %, 15. 94 % and 30. 77 %, respectively, and that of group 3, 2 was 4 and 2 times as much as that of group 1, respectively (OR = 4. 00, P<0. 001; OR=2. 343, P<005, respectively). It was concluded that HCMV-IgM(+) can only be considered as an screening indicator for pregnant women with HCMV infection, while IgM(+) combined with serous DNA( + ) or mRNA( + ) indicates active infection and has a high incidence of maternal-fetal transmission and abnormal pregnancy outcome.

关键词： human cytomegalovirus enzyme linked immunosorbent assay polymerase chain reaction : pregnancvaction pregnancy

A NEW QUANTITATIVE DETECTION METHOD OF RECOMBINANT CFP10-ESAT6 AMALGAMATION PROTEINS FROM MYCOBACTERIUM TUBERCULOSIS BASED ON MICRO-MAGNETIC PROBES STRATEGY

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Journal of Innovative Optical Health Sciences 2012年第1期5卷 55-60页

作者： YIQING HUANG JINPING LUO MIXIA WANG JUNTAO LIU XINXIA CAI State Key Laboratory of Transducer Technology Institute of Electronics Chinese Academy of Sciences Beijing 100190China Graduate School of Chinese Academy of Sciences Beijing 100190 China

A new rapid,specific and sensitive method for assay of recombinant CFP10-ESAT6 amalgamation proteins from Mycobacterium tuberculosis was proposed.The method used streptavidincoated magnetic beads to enrich the specific biotinylated anti-CFP10 antibody,then adopted a sandwich-type enzyme linked immunosorbent assay technology with two kinds of monoclonal antibodies:biotinylated anti-CFP10 antibody and HRP-labeled anti-CFP10 antibody to identify the target CFP10-ESAT6 proteins,and finally detected chemiluminescence intensity by a small home-made optical sensor.It was shown that,the corresponding chemiluminescence intensity had a good logarithmic linear response to the concentration of CFP10-ESAT6 proteins when ranging at 1~1000 ng/mL,and the correlation coefficient is 0.9937.The proposed method could detect the CFP10-ESAT6 proteins with low detection limit(1 ng/mL)and the detection time could be controlled within 45 min.Compared with commonly used detection methods of M.tuberculosis,this method was easy to operate,faster,and of higher sensitivity.The achievement of the quantitative detection of CFP10-ESAT6 proteins has important scientific significance and wide application prospects in tuberculosis control.

关键词： enzyme linked immunosorbent assay chemiluminescence home-made optical sensor

Association of Preoperative Serum IGF-Ⅰ Concentration with Clinicopathological Parameters in Patients with Non-Small Cell Lung Cancer

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2013年第2期33卷 224-227页

作者：付圣灵唐和孝廖永德江文洋徐沁孜邓豫付向宁 Department of Thoracic Surgery Tongji Hospital Tongji Medical College Huazhong University of Science and Technology

Insulin-like growth factor- I （IGF- I ） is a mitogenic and anti-apoptotic factor. Serum IGF-I concentration is related to some cancer risk and tumor progression. The aim of this research was to study the association of preoperative serum IGF- I concentration with clinicopathological parameters and prognosis of non-small cell lung cancer （NSCLC）. Preoperative serum IGF- I concentration was measured in 80 consecutive patients with NSCLC who underwent radical lung cancer resection, and 45 patients with benign pulmonary lesion （BPL） by Using enzyme linked immunosorbent assay （ELISA）. The results showed that the serum IGF- I concentration was elevated and correlated with clinicopa- thological parameters and overall survival （OS） in NSCLC patients. Serum IGF- I concentration was significantly higher in patients with NSCLC tfian in those with BPL. The IGF- I concentrations were significantly higher in NSCLC patients with 〉T2, NI-3, and in IIIA-IV but not in those with 〈T2, NO, or I A- IIB. The increased serum IGF- I concentration was significantly correlated with poor prog- nosis. Our data show the positive correlation between IGF-I serum concentration and the tumor size for the first time. It seems that IGF-I related to the progression of lung cancer may depend on autocrine/paracrine function. In addition, our study reveals that higher serum IGF- I concentration is correlated with larger tumor size, advanced stages, local lymph node metastasis and worse prognosis, indicating that endocrine IGF- I is also important for the progression for NSCLC.

关键词： non-small cell lung cancer insulin-like growth factor I enzyme linked immunosorbent assay prognosis

DETERMINATION OF SERUM SOLUBLE MACROPHAGE COLONY-STIMULATING FACTOR RECEPTOR LEVELS IN PATIENTS WITH HEMATOLOGICAL DISEASES

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Chinese Journal of Cancer Research 2001年第3期13卷 185-189页

作者：饶青韩敬淑沙晓津杨仁池耿以琪郑国光吴克复 Institute of Hematology & Blood Diseases Hospital Chinese Academy of Medical Sciences & Peking Union Medical College Tianjin

Objective: To investigate the serum levels of soluble macrophage colony-stimulating factor receptor (M-CSFsR) in normal subjects and patients with hematological diseases and its clinical implications in hematological diseases. Methods: The concentration of M-CSFsR was determined by ELISA. The serum M-CSFsR was identified and characterized by immunoprecipitation and Western blotting. Results: The mean serum level of M-CSFsR of 123 normal individuals was 0.48 ng/ml ± 0.41 ng/ml. Immunoprecipitation and Western blotting assay revealed a ～ 90kD band of serum M-CSFsR. The mean serum M-CSFsR level of 60 patients with acute lymphoblastic leukemia (ALL), 36 patients with acute myeloblastic leukemia (AML), 13 patients with myelodysplastic syndrome (MDS) and 42 patients with aplastic anemia (AA) .were 0.22 ng/ml±0.23 ng/ml, 0.17 ng/ml±0.16 ng/ml, 0.19 ng/ml±0.16 ng/ml and 0.23 ng/ml±0.21 ng/ml, respectively, which were significantly lower than that of normal subjects (P=0.002 ,P<0.0001,P<0.0001 andP<0.0001). The mean serum M-CSFsR level of 51 idiopathic thrombocytopenic purpura (ITP) patients was significantly higher than that of normal subjects (2.05 ng/ml±2.75 ng/ml,P<0.0001). Conclusion: The serum M-CSFsR levels of patients with ALL, AML, MDS and AA were significantly lower, while the level of patients with ITP was significantly higher than that of normal individuals. Patients with severe ITP (platelet count<30×l09/L) had the highest M-CSFsR level. It suggested that the abnormal levels of serum M-CSFsR may associate with some hematological diseases and may contribute to the pathological process.

关键词： Macrophage colony-stimulating factor Receptor enzyme linked immunosorbent assay Immunoprecipitation Western blotting Leukemia Idiopathic thrombocytopenic purpura

The relationship between the levels of granulocyte colony stimulating factor and leukocytosis induced by all trans retinoic acid in acute promyelocytic leukemia

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Chinese Medical Journal 1999年第12期 29-31页

作者：吴文孙关林邬维礼周荣富李秀松沈志祥王振义

Objective To explore the mechanism of leukocytosis Methods enzyme linked immunosorbent assay (ELISA) method was used for detecting levels of serum granulocyte colony stimulating factor (G CSF) in 47 cases of acute promyelocytic leukemia (APL) during the treatment with all trans retinoic acid (ATRA) Results The peak of increased serum G CSF level occurred on the 9th day, and WBC number was the highest on the 11th day After ATRA treatment, both serum G CSF level and WBC number increased in 68 1% of the cases In 19 2% of the cases treated, serum G CSF level was increased but without obvious change in WBC number, and the reverse was true in 12 7% of the cases Conclusion Serum G CSF level was statistically correlated to the number of WBC, promyelocytes and its late stage by Spearman's rank order correlation coefficient

关键词： leukocytosis enzyme linked immunosorbent assay granulocyte colony stimulating factor retinoic acid leukemia

Development of PPA-ELISA for Detecting Antibodies against Porcine Pseudorabies Virus Using Truncated Recombinant Glycoprotein gD Expressed in E.coli

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Animal Husbandry and Feed Science 2011年第6期3卷 29-34页

作者： ZU Li-chuang SHEN Zhi-qiang LI Jiao WANG Jin-liang Shandong Lvdu Biological Technology Co. Ltd. Binzhou 256600 China Shandong Binzhou Animal Science & Veterinary MedicineAcademy Binzhou 256600 China

The purpose of this study was to develop a method for detecting antibodies against porcine pseudorabies virus （PRV）. According to the published genomic sequence of PRV SA strain, an approximately 1 070-bp gD gene fragment was amplified by PCR. The PCR products were cloned into the prokaryotic expression vector pET30a and the positive recombinant plasmid was transformed into E. coli BL21. Through induction with IPTG, the recombinant gD protein was expressed as inclusion bodies. As analyzed by western blot assay, the purified recombinant gD protein had good antigenicity and high specificity. Using the purified gD protein as coating antigen and horseradish peroxidase labeled staphylococcal protein A （PPA） as secondary antibody, we developed a PPA-ELISA for detecting antibodies against porcine PPV. No cross-reaction with the positive sera against seven common pathogens in pigs including classical swine fever virus, porcine parvovirus, porcine reproductive and respiratory syndrome, Japanese encephalitis virus, porcine circovirus type 2, porcine epidemic diarrhea virus, transmissible gastroenteritis virus was observed. The repeatability test showed that the intra- and inter-assay coefficients of variation were lower than 5% and 10%, respectively. Compared with the ELISA gD antibody test kit produced by IDEXX, the coincidence, sensitivity and specificity of the developed PPA-ELISA were 92.0%, 95.1% and 88.1%, respectively. The developed PPA-ELISA had good repeatability, sensitivity and specificity and was a rapid and simple serological method for surveillance of PRV antibodies in pig herds as well as for rapid diagnosis and epidemiological investigation of PRV infection.

关键词： Porcine pseudorabies virus gD protein Truncated expression enzyme linked immunosorbent assay

Development of an Indirect ELISA Using Recombinant Truncated Envelope Glycoprotein for Detection of Antibodies against Japanese Encephalitis Virus

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Animal Husbandry and Feed Science 2010年第1期2卷 38-42页

作者： ZU Li-chuang WANG Jin-liang GUAN Yu SHEN Zhi-qiang DONG Lin LI Jiao Shandong Ludu Bio-Technology Co. Ltd. Binzhou 256600 China Shandong Binzhou Animal Science and Veterinary Medicine Academy Binzhou 256600 China

[ Objective] To develop an indirect ELISA assay for detecting antibodies against envelope glycoprotein （ E protein） of Japanese encephalitis virus （JEV）. [ Method] Specific primers were designed according to JEV sequences published in the GenBank. The cDNA of JEV E gene （about 1 000 10p） was amplified by the RT-PCR with the specific primers. After sequencing analysis, the E gene was cloned into pET30a expression vector and expressed in E. coli BL21 （DE3） with the induction of IPTG. After denaturation, purification and renaturation, the recombinant protein was analyzed by the SDS-PAGE and the westem blotting. An indirect ELISA was developed to detect antibodies against JEV. [ Result] The E protein was mainly expressed in inclusion body. With the purified E protein, the indirect ELISA was developed and displayed good specificity, sensitivity and repeatability, [ Conclusion]The developed ELISA using the truncated E protein as antigen is a simple, convenient and rapid serological method for diagnosis, monitoring antibody level and epidemiological investigation of JEV.

关键词： Japanese encephalitis virus E protein Prokaryotic expression enzyme linked immunosorbent assay Antibodies

Evaluation of Usefulness of Three Serological Tests Using Native Crude Antigen in Diagnosis of Hepatic Cystic Echinococcosis Patients

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Open Journal of Medical Microbiology 2021年第2期11卷 69-79页

作者： Mesut Akil Ahmet Ozkeklikci Eylem Akdur Ozturk Aygul Sadiqova Nuray Altintas Selda Karamil Ozge Sarica Yilmaz Aysegul Unver Nazmiye Altintas Department of Parasitology Faculty of Medicine Ege University Izmir Turkey Department of Microbiology Dr. Ersin Arslan State Hospital Gaziantep Turkey Department of Medical Biology Faculty of Medicine Celal Bayar University Manisa Turkey

Objective: To evaluate three different serological tests [Indirect Hemaglutination (IHA), enzyme linked immunosorbent assay (ELISA) and Western Blotting (WB)] using native crude antigen for diagnosis of hepatic cystic echinococcosis (HCE) patients. Materials and Methods: Sheep hydatid fluid (HF) was collected from fertile cysts obtained from a slaughterhouse and used as an antigen. Forty patients who were attended the Dr. Ersin Arslan Training and Research Hospital in Gaziantep, Turkey, were investigated. Serum samples were obtained from surgically confirmed CE patients. Healthy Turkish people and 16 patients with other helminthic infections were included as a control group. Results: Of the 40 analyzed patients, 10 (25%) were men and 30 (75%) were female. The average age was 46.97 years (s.d.;18.95). The majority of the patients had a single cystic lesion situated in one lobe of the liver (usually in the right lobe) (55%), 32.5% of patients had two cystic lesions and 12.5% of patients had multiple cyst formations with various numbers. In all cases, ultrasound (US) examinations were positive and the size of cysts was between 2.1 - 12.7 cm. Twenty-three patients of the total 40 patients were classified according to the WHO classification system based on US findings. According to the results of WB analysis, molecular weights of 8 kDa (80%), 12 kDa (80%), 22 - 24 kDa (97.5%), 26 kDa (97.5%), 34 kDa (100%), 36 - 38 kDa (90%), 45 - 50 - 55 kDa (97.5%), and 60 - 75 kDa (97.5%) bands were identified. But 34, 50, and 55 kDa bands were also found in other helminthic diseases. Conclusion: The specificity and sensitivity of three serological tests (IHA, ELISA and WB) using crude antigen were compared by diagnosing hepatic cystic echinococcosis patients. IHA and ELISA showed high sensitivity but low specificity. Western blotting showed low sensitivity but high specificity.

关键词： Cystic Echinococcosis Serodiagnosis enzyme linked immunosorbent assay Indirect Hemagglutination assay Western Blotting