Development of a class-specific polyclonal antibody-based indirect competitive ELISA for detecting fluoroquinolone residues in milk

Modified 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) method was employed to synthesize the artificial antigen of norfloxacin (NOR), and New Zealand rabbits were used to produce anti-NOR polyclonal antibody (pAb). Based on the checkerboard titration, an indirect competitive enzyme-linked immunosorbent assay (icELISA) standard curve was established. This assay was sensitive and had a working range from 0.12 to 68.40 ng/ml, with the half maximal inhibitory concentration (IC₅₀) and limit of detection (LOD) values of 2.7 ng/ml and 0.06 ng/ml, respectively. The produced pAb exhibited high cross-reactivity to fluoroquinolones (FQs) tested, and the IC₅₀ values to enoxacin, ciprofloxacin, and pefloxacin were 3.1, 3.4, and 4.1 ng/ml, respectively. It also indicated that the concentrations of NaOH and methanol in assay buffer should not be higher than 10% and 30%. When spiked in milk at 5, 20, and 50 ng/ml, the recoveries for NOR, enoxacin, ciprofloxacin, and pefloxacin ranged 90.5%–98.0%, 84.0%–95.2%, 94.0%–106.0%, and 89.5%–100.0%, respectively. The results suggest that this class-specific pAb-based icELISA could be utilized for the primary screening of FQ residues in animal-original products.     

“即刻性”质控的应用

借助质控血清开展室内质控,可以控制实验的重复性、精密度,从而使检测结果趋于稳定．目前,ＥＬＩＳＡ免疫实验室仍采用临床化学测定使用的质控方法开展室内质控,由于受ＥＬＩＳＡ方法特殊性限制（试剂盒有效期短,有些项目测定次数少等）,此质控方法在具体操作中有其缺陷性,我室在此质控方法基础上,同时进行“即刻性”质控（只需连续检测３次,即可对第３次结果进行质控）．有效弥补了上述不足,取得了很好的质控效果．     

阳泉市中小学生HBsAg检测结果分析

目的:了解阳泉市中小学生HBsAg携带情况,为制定防治措施提供依据.方法:采集被监测者血用酶联免疫吸附实验做HBsAg检测,结果用Excel统计分析.结果:监测1986名学生HBsAg阳性者43名,总阳性率为2.16%;其中,私立学校阳性率为2.31%,公办学校阳性率2.05%;男生阳性率为2.76%,女生阳性率1.39%;私立学校小学、初中、高中生HBsAg阳性率分别为1.44%、2.26%、2.87%,有随年龄增长而增加的特点.结论:阳泉市中小学生HBsAg阳性率明显低于全国总体水平,但HBsAg防治工作不能放松,要加强对住校生的管理工作,对易感者普遍接种乙肝疫苗.     

冠心病与肺炎衣原体感染的相关性研究

目的：测定冠心病患者肺炎衣原体（CPn）感染情况，并与健康对照组比较．探讨CPn感染和冠心病的关系。方法：采集100例冠心病（CHD）患者空腹静脉血，健康对照者60例，采用ELISA法对血浆中CPn IgG进行检测。结果：100例冠心病人血浆CPn IgG阳性51例，占51．00％；60例健康对照组血浆CPnIgG阳性20例．占33．33％。冠心病组CPn IgG阳性率高于健康对照组，经X^2检验，X^2=4．472，P=0．029，两组肺炎衣原体感染比较有统计学意义。结论：冠心病组血浆CPn IgG阳性率高于健康对照组，肺炎衣原体在冠心病的发病过程中起到一定的作用。     

Development of an indirect competitive ELISA for simultaneous detection of enrofloxacin and ciprofloxacin

Modified 1-ethyl-3-(3-dimethylaminopropy) carbodiimide (EDC) method was employed to synthesize the artificial antigen of enrofloxacin (ENR), and New Zealand rabbits were used to produce anti-ENR polyclonal antibody (pAb). Based on the checkerboard titration, an indirect competitive enzyme-linked immunosorbent assay (ELISA) standard curve was established. This assay was sensitive and had a linear range from 0.6 to 148.0 μg/kg (R ²=0.9567), with the half maximal inhibitory concentration (IC₅₀) and limit of detection (LOD) values of 9.4 μg/kg and 0.2 μg/kg, respectively. Of all the competitive analogues, the produced pAb exhibited a high cross-reactivity to ciprofloxacin (CIP) (87%), the main metabolite of ENR in tissues. After optimization, the matrix effects can be ignored using a 10-fold dilution in beef and 20-fold dilution in pork. The overall recoveries and coefficients of variation (CVs) were in the ranges of 86%–109% and 6.8%–13.1%, respectively. It can be concluded that the established ELISA method is suitable for simultaneous detection of ENR and CIP in animal tissues.     

Immunodiagnosis of tuberculosis: An update

Tuberculosis is still a major health problem in most developing countries and its incidence is rising in many developed countries. This resurgence has been attributed to the HIV epidemic and TB has been declared as a global health emergency by WHO in 1993. The diagnosis of tuberculosis mainly depends upon initial clinical suspicion and radiographic findings with subsequent bacteriological confirmation by sputum smear examination and culture. Lack of sensitivity in smear examination, non specificity of radiological findings, extended tum around time ofMycobacterium tuberculosis culture and difficulties in diagnosing paucibacillary, childhood and extrapulmonary tuberculosis has necessitated to explore the utility of immunodiagnosis of tuberculosis as a convenient and cost effective test to supplement clinical information for definite diagnosis. Many commercial tests are available in the market for diagnosis of TB. Most of these tests are based on the detection of IgG, IgA and IgM antibodies to specific mycobacterial antigen or mixture of antigens. Indigenous immunoassay systems have explored excretory-secretory ES-31 mycobacterial antigen for immunodiagnosis of TB. Many a time there is lack of consistent elevation in all the three Ig classes in active infection thus making it more important to determine the ideal antibody isotype assay for reliable diagnosis of tuberculosis and to save the costs of the patient for unnecessary investigations.     

Serodiagnosis of tuberculosis using two ELISA systems

Serodiagnosis by ELISA has been widely explored over the years, in the diagnosis of tuberculosis. Two ELISA systems were evaluated for detection of mycobacterial antibodies in pulmonary and extra pulmonary tuberculosis. The two test assays explored were ERBA LISA (TB IgG) test (Anda Biologicals) which uses A60 antigen complex found in the cytosol of typical and atypical mycobacteria, and SEVA TB (IgG) ELISA, which uses a 31 kDa, glycoprotein antigen purified fromM. tb H₃₇Ra culture filtrate. Sera from 98 proven tuberculosis [pulmonary TB (48), tuberculous lymphadenopathy (30), tuberculous meningitis (15) & genitourinary TB (5)] were studied along with 32 healthy controls. The overall positivity obtained using ERBA LISA (TB IgG) test and SEVA TB (IgG) ELISA test was 72.9% and 91.6% in pulmonary tuberculosis, 43.3% and 76.6% in tuberculous lymphadenopathy respectively. The sensitivity of ERBA LISA test in tuberculous meningitis and genito-urinary TB was significantly low (26.6% & 40% respectively) compared to sensitivity obtained using SEVA TB ELISA (86.6% & 60% respectively) with overall specificity of 60% and 87.5%. Thus SEVA TB IgG ELISA test was found to be more sensitive than ERBA LISA in detecting IgG antibodies in tuberculous sera, in particular in extra pulmonary tuberculosis cases.     

光敏感核不育水稻花粉败育过程中花药的差示扫描量热法(DSC)研究

本文以湖北光敏核不育水稻(Hubei Photoperiod Sensitive Genio Male—Sterile Rice i·e·HPGMR)农垦58A和常规品种农垦58为材料,在人工控制的长,短日照条件下,用DuPont9900型差示扫描量热计测取了不同发育时期花药的DSC谱。结果表明:可育花药的DSC谱上的低温吸热峰面积随发育呈现周期性变化;不育花药在单核中期以前与可育花药差别不大,而在单核中期以后的DSC谱上的低温吸热峰面积明显小于可育花药,且不呈现周期性变化,而逐渐减小到接近对零。     

Age-Related Changes of Serum Soluble Interleukin 9 Receptor (sIL-9Rα) in Healthy Subjects

Most cytokine receptors including interleukin (IL)-9 have soluble counterparts in body fluids. We planned to investigate the pathophysiological significance of the serum soluble IL-9 receptor (sIL-9R) level. We determined the serum sIL-9Rα chain (sIL-9Rα) levels in 96 healthy Japanese individuals to establish a control value by means of specific human sIL-9Rα ELISA, followed by a preliminary application in a patient with diarrhea positive hemolytic uremic syndrome. Age was negatively correlated with the sIL-9Rα level (Spearman r = −0.241, n = 96, p = 0.0180). The serum sIL-9Rα level showed a progressive decline to the normal adult level by the age of 30. The serum sIL-9Rα level of the patient with HUS was markedly higher than those of the age-matched control from the onset of the disease. Because of the remarkable age-dependent variability of sIL-9Rα in healthy subjects, disease-related changes, as well as therapy-dependent alterations, should be considered with caution. Thus, it is recommended that when the serum sIL-9Rα levels of patients are evaluated, the values should be compared with those of age-matched controls. The established control value will be used to discriminate between normal and the pathological conditions in our future studies.     

Monoclonal antibody-based ELISA and colloidal gold-based immunochromatographic assay for streptomycin residue detection in milk and swine urine

A protein conjugate of streptomycin (streptomycin-bovine serum albumin (BSA) conjugate) was prepared and used as immunogen to produce monoclonal antibodies (MAb). One hybridoma secreting anti-streptomycin MAb was obtained and then used to produce MAb. The MAb named 13H5 showed the 50% maximal inhibitory concentration (IC50) value of 4.65 ng/ml and the IC20 value of 0.21 ng/ml in phosphate buffered saline (PBS). At optimum conditions, an indirect competitive enzyme-linked immunosorbent assay (ELISA) and a co...