本文根据2016年实施的《JJG1125-2016氯乙烯气体检测报警仪》计量检定规程,评定了氯乙烯气体检测报警器示值误差测量结果的不确定度,与计量同行交流.

原子荧光法检测环境样品汞、砷、硒,从采样到化验室检测过程中砷、硒都非常稳定,受外界干扰很小,但由于汞具有蓄积性,某一环节稍不注意就会影响检测结果.本文从采样到实验室各个环节分别阐述了对实验结果有影响的各个因素,以保证实验检测结果的准确性.

建立了一种合理科学的挥发性有机物氢火焰离子化检测仪的计量校准方法.根据仪器的工作原理和使用要求,提出了示值误差、响应时间、重复性、仪器稳定性等校准项目,并对示值误差的测量结果不确定度进行了评定.通过实例分析表明,该方法可以有力的保证仪器的溯源性和实际检测中的准确性.

目的：探讨他克莫司药物浓度检测过程中数据异常原因及其相应的解决对策。方法对80例患儿入院资料进行分析,患儿入院后对其进行常规检查,如：血压、肝肾功能等,并对患儿进行血药浓度监测,分析他克莫司药物浓度检测过程中数据异常原因及其针对性的解决措施。结果所有患儿均在1个月内和1~5个月内进行药物浓度检测,1个月内他克莫司药物浓度正常者占28.5%,17.5%患儿低于参考值,少于1~5个月内检测浓度,差异具有统计学意义(P<0.05)；1个月内53.8%患儿他克莫司药物浓度高于参考值,多于1~5个月内检测浓度,差异具有统计学意义(P<0.05)。结论他克莫司药物浓度检测过程中数据异常影响因素较多,因此患儿检测过程中必须对检测结果提供合理解释,并根据异常原因提出相应的应对措施,保证患儿能够正确给药,提高临床治疗水平。

Objective To determine the prevalence of antibodies to cyclic citrullinated peptides (antiCCP) in patients with juvenile-onset systemic lupus erythematosus (JSLE) and its potential clinical significance. Methods Anti-CCP was measured in sera from patients with JSLE (n=47), juvenile idiopathic arthritis (JIA, n=54) and the sera from age-matched healthy children (n=40) using the third generation of anti-CCP ELISA commercial kit. The association of anti-CCP with other laboratory parameters and clinical features, especially arthritic symptoms in JSLE was also analyzed. T-test, Mann-Whitney U test, Chi-square and Fisher's exact test were used for statistical analysis. Results Out of the 47 JSLE patients, 6 (13%) were anti-CCP positive, which was significantly higher than that of the healthy controls( 13% vs 0, P＜0.05 ), but not different from that of the JIA group (26%, P=0.098). RF was more prevalent in JSLE patients with anti-CCP than patients without (83% vs 15%, P＜0.01 ), but there was no difference in other laboratory parameters and the clinical features ineluding the occurrence of arthritis (67% vs 51%, P＞0.05). As one of the initial symptoms, arthritis was observed in 25 of 47 JSLE patients and no one had developed deforming arthropathy.There was no statistical difference in anti-CCP positivity between JSLE patients with and without articular involvement ( 16% vs 9%, P＞0.05 ). Anti-CCP was not detected in any of the 3 patients with JSLE who had experienced joint pain and limited activity during 3 years follow-up. Conclusion Anti-CCP could be detected in patients with JSLE. It is noteworthy when differentiate from juvenile idiopathic arthritis, but the presence of anti-CCP does not relate with the occurrence of arthritis at presentation and persistence of arthritis in JSLE.

Objective To conduct the amplification,cloning,bioinformatics analysis,prokaryotic expression and purification of enterovirus 71 VP1 gene segment and to initially confirm the biological activity of the recombinant expression product.Methods A pair of specific primers was designed according to GenBank EV71 sequence,viral RNA as a template was extracted from the throat swab specimens in the EV71 patients.EV71 VP1 gene was amplified by RT-PCR.After enzyme digestion,the expression vector pET28a was inserted.The prokaryotic expression vector of pET28a-EV71 VP1 was constructed.Then the E.coli DH5a transforma-tion was performed.IPTG was adopted for induction expression.The expression results were analyzed by using SDS-PAGE and Western blot.The bioinformatics analysis of the sequenced results was performed by the software.Expressed protein was purified and the plates were coated,ELISA was used to test the VP1 specific IgG antibody in serum samples of EV71 positive and COX A16-positive patients.Results The BLAST alignment showed that the homology of the objective gene EV71 VP1 was 99% com-pared with other strains(JQ766207.1)in GenBank.EV71 VP1 protein was about 32×103 ,which mainly existed in the form of in-clusion body.The bioinformatics analysis showed that EV71 VP1 protein was a hydrophilic protein,without transmembrane region and N-terminal signal peptide sequence,the tertiary structure existed.The ELISA results showed that the specific IgG OD value in EV71-positive patients was(2.425±0.521),OD value in COX A16 positive patients was(1.205 ±0.314),the normal control OD value was(0.353±0.128).The sensitivity and specificity of EV71 VP1 protein detection were 84% and 88% respectively.Conclu-sion The pET28a-EV71 VP1 expression vector is successfully constructed;the preliminary analysis on the serum of the infected patients by ELISA shows that the obtained objective protein has higher sensitivity and specificity,which is initially confirmed to have biological activity and can be further used for the related study on EV71 diagnosis and vaccine.

Objective To investigate the significance of IL-18in diagnosis of macrophage activation syndrome(MAS)due to systemic-onset juvenile idiopathic arthritis(SoJIA).Methods Concentrations of IL-18were quantified in serum of the four groups(patients with MAS complicating SoJIA,patients with SoJIA,patients with JIA but not SoJIA and controls)by ELISA.Results Serum IL-18concentrations of MAS complicating SoJIA patients are significantly higher than those of any other patients in the study,SoJIA patients are the second high in the four groups,and the IL-18concentrations in controls are the lowest.Conclusion IL-18may be important in the diagnosis of MAS complicating SoJIA.

Objective To understand the age distribution of children allergic to cockroach,and diseases sources in Guangzhou.Methods Cockroach antigen specific IgE was detected by modified RAST assay.The positive results of January 2010 to December 2012 were analyzed.Results Among 9 821 serum samples,245 cases were positive for cockroach antigen specific IgE detection,with a positive rate of 2.49%.The top five symptoms in children positive for cockroach specific IgE were urticaria (59 cases,30.10%),allergic rhinitis(45 cases,22.96%),asthma (35 cases,17.86%),Henoch-Schonlein Purpura(21 cases,10.71%),and conjunctivitis (11 cases,5.61%).The age distribution of these patients was mainly concentrated in the 4 to 10 years of age,of which 10.57% was 4 years old,13.82% was 5 years old,11.38% was 6 years old,12.20% was 7 years old,and 10.98% was 8 years old.Male to female ratio was 2.77∶1.Conclusions Cockroach allergen was one of the major allergens for children in Guangzhou.Allergies were occurs primarily in children with allergic rhinitis and urticaria.The age distribution of patients was mainly in 4 to 10 years old.The positive rate of male was higher than that of female.

Objective To investigate the value of antikeratin antibody(AKA),anti-cyclic citrullinated peptide antibody(anti-CCP)and rheumatoid factor(RF-IgG,RF-IgM)in early diagnosis and prediction of juvenile idiopathic arthritis(JIA).Methods Seventy-six patients with JIA were enrolled and follow-up at least 6 months.The blood samples were collected to measure AKA,anti-CCP,RF-IgG,RF-IgM upon admission.The frequency of positive AKA,anti-CCP,RF-IgG and RF-IgM in different subtype and refractoriness were compared.Forty-nine healthy children were enrolled as control group.Results Compared with control group,the positive rates of AKA,anti-CCP,RF-IgG and RF-IgM were significantly higher in polyarticular subgroup,but there were no difference in other subtypes.The sensitivity was AKA > anti-CCP,RF-IgG > RF-IgM.The specificity was RF-IgM > AKA > RF-IgG > anti-CCP.The positive rate of AKA was significantly higher in refractory JIA than that in common JIA.The relative risk estimate(OR)was 3.514.Conclusions AKA,anti-CCP,RF-IgG,RF-IgM have different values in early diagnosis and the subtype of JIA.AKA,anti-CCP are sensitive and specific in distinguishing polyarticular subgroup.AKA may predict refractory JIA.

Objective To investigate the value of antikeratin antibody(AKA),anti-cyclic citrullinated peptide antibody(anti-CCP)and rheumatoid factor(RF-IgG,RF-IgM)in early diagnosis and prediction of juvenile idiopathic arthritis(JIA).Methods Seventy-six patients with JIA were enrolled and follow-up at least 6 months.The blood samples were collected to measure AKA,anti-CCP,RF-IgG,RF-IgM upon admission.The frequency of positive AKA,anti-CCP,RF-IgG and RF-IgM in different subtype and refractoriness were compared.Forty-nine healthy children were enrolled as control group.Results Compared with control group,the positive rates of AKA,anti-CCP,RF-IgG and RF-IgM were significantly higher in polyarticular subgroup,but there were no difference in other subtypes.The sensitivity was AKA anti-CCP,RF-IgG RF-IgM.The specificity was RF-IgM AKA RF-IgG anti-CCP.The positive rate of AKA was significantly higher in refractory JIA than that in common JIA.The relative risk estimate(OR)was 3.514.Conclusions AKA,anti-CCP,RF-IgG,RF-IgM have different values in early diagnosis and the subtype of JIA.AKA,anti-CCP are sensitive and specific in distinguishing polyarticular subgroup.AKA may predict refractory JIA.

目的 初步了解抗中心体抗体(ACA)在儿童病原感染及自身免疫性疾病中的临床意义.方法 收集广州市儿童医院2009年4月至9月19例ACA检测阳性患儿为ACA阳性组、42例ACA阴性为阴性对照组,选取同期常见病原微生物感染及正常儿童为阳性对照组和正常对照组.对4组病原微生物感染指标和其他自身抗体及炎症相关蛋白资料进行统计学分析.结果 ACA阳性存在于自身免疫相关疾病如系统性红斑狼疮(SLE)等16例次(84.2%);感染因素相关疾病7例次(36.8%).ACA阳性组与阴性对照组间病原微生物感染指标差异无统计学意义(P>0.05);两组间ANA阳性率差异有统计学意义(P<0.001).阳性对照组、正常对照组与ACA阳性组间病原微生物感染指标差异无统计学意义(P>0.05).结论 儿童ACA常见于免疫性疾病如SLE、川崎病,与机体亢进的自身免疫状态有关,而与以往报道的MP、CP、HSV等病原微生物感染无直接关系.

近十年来,RA实验室诊断取得了引人瞩目的发展,ACPAs作为一类针对瓜氨酸蛋白/肽的高特异性且灵敏的血清学标志,已被广泛应用于临床[1].虽然JIA患儿的关节病变与成人RA相似,但JIA极度缺乏特异性诊断指标,尽管国内外儿科风湿病学者对可否将ACPAs引入JIA诊断和预后中显示出浓厚的兴趣,然而结果与期望相去甚远[2-8].目前的研究主要集中于IgG类抗-CCP,其他ACPAs鲜见报道.本研究采用同时检测抗-CCP IgG/IgA以及CCP之外的靶抗原等5种不同ELISA商品试剂盒检测一组JIA患儿ACPAs,以探讨其在JIA诊断中的价值与临床意义.

Objective To investigate the distribution of common allergens and cross reaction of house dust mites and dust mites in children with allergic diseases in Guangzhou area.Methods Total 1136 children with allergic diseases(298 infants,455 preschool children and 403 school-age children)were recruited from July 2009 to July 2010.Thirty-five kinds of serum specific-IgE were detected by MAST system.The positive rates of each allergen among three age groups were compared.Meanwhile the cross reaction of house dust mites and dust mites were analyzed.Results Among the 1136 children,the top three allergens were dust mites 774(68.13%),house dust 557(49.03%),and milk 411(36.18%).The top three allergens in infants were milk(195,70.14%),cheese(133,47.84%),and egg(102,36.69%).The top three allergens in preschool children were dust mite(371,80.13%),dust(311,68.35%),and milk(177,38.90%).The top three allergens in school age children were dust mite(306,75.93%),dust(163,40.45%)and animal hair(104,25.81%).In 774 children allergic to dust mite,685(88.50%)showed positive reaction to both Dermatophagides pteronyssinus(Der p)and Dermatophagides farina(Der f).9.56% showed positive reaction to Der f and 1.94% to Der p.There were no statistical differences of positive reaction to dust mites among three age groups(P 0.05).Conclusions The distribution of common allergens in allergic children in Guangzhou has been changes to some extent and is also some what different from other areas in China.A considerable proportion of allergic children have positive reaction to both Der p and Der f.Only a few of allergic children have positive reaction to one of dust mites.It merits attention in clinical desensitization treatment.

Objective: To evaluate the clinical diagnostic value of anti-RA33 in patients with Juvenile idiopathic arthritis(JIA).Methods: The level of serum Anti-RA33 antibody and rheumatoid factor(RF) were detected by Enzyme linked immunoabsorbent assay(ELISA) in serum samples from 81 cases with JIA(including 19 female,62 male,with average age of 8.6 years,with an average history of 1.4 years),while 55 cases with SLE,other joint diseases or virus infections and 49 normal children were enrolled to serve as control.The positive and negative results are diagnosed by critical value of kit.Results: The positive ratio of anti-RA33 and RF in 81 JIA cases were 11.11%(9/81) and 12.35%(10/81),respectively,compared with nomal control,has statistical significance(P0.05).But the positive ratio of anti-RA33 and RF has no significant different between the JIA group and other joint disease group(P0.05).There is no dependability between RF and the detection of anti-RA33 in JIA group(P0.05).In all kinds of JIA hypotype,anti-RA33 antibody is principally reside in systerm arthritis and polyarthritis,respectively,33.3% and 25.0%,which has significantly difference(P0.05).RF only appears in polyarthritis,with 62.5 percent.18 cases have imaging changes in those 81 JIA cases and 4 cases of which has positive anti-RA33 antibody(22.2%).Compared to JIA chidren without imaging changes,there is no significant difference(P0.05).Conclusion: Anti-RA33 antibody still can not be recognized as new and reliability index to diagnostic efficiency in children with early JIA.Anti-RA33 are principally reside in systerm arthritis and polyarthritis,which may provide guidance on the classification of JIA.

目的 建立广州地区1～8岁儿童血清补体C3,C4含量的参考范围.方法 应用Beckman-coulter Immage 800特种蛋白分析仪,采用免疫比浊法对968例1～8岁健康儿童血清补体C3,C4含量进行检测,分成7个年龄段对结果进行统计分析.结果 广州地区1～8岁健康儿童血清补体C3,C4含量在性别和各年龄段之间差别无统计学意义(P>0.05),总的参考范围(g/L)为C3:0.59～1.81,C4:0.19～0.22.同时与全国临床检验操作规程以及厂家提供的参考范围相比较差异均有统计学意义(P<0.05).结论 1～8岁儿童血清补体C3,C4旧的参考范围已不适用于当今人群,各地、各实验室应测定一批健康人的标本,统计出各自实验室的参考范围,若存在性别差异,应分别制定两性的参考范围,若存在年龄组间差异,还应进一步制定两性间各个年龄组的参考范围.

Objective To determine the basal level of serum immunoglobulin of 1 to 8 years old children. Methods 968 children between 1 to 8 years old were included in this study. Levels of serum immunoglobulin were determined by the Beckman-coulter Immage 800 protein analyer and the data were analyzed with statistical methods. Results Variation in the level of serum immunoglobulin was observed in children of differ sex and age. The results were also differed from the national values and the values provided by the manufacturer. Conclusion The values obtained from this group of children are not applicable to other children. Further analysis on the level of serum immunoglobulin from the normal subject from other laboratories is required.

目的 调查乙型肝炎表面抗体(抗HBs)定性检测情况,探讨影响实验室结果差异的因素.方法 通过实地或电话方式采访检验科免疫室负责人或实验操作人员,随机抽取广州市二级以上33所医疗机构,就抗HBs实际检测情况进行调查.结果 所凋查的医疗机构在结果判读上存在严重主观性,33所机构中有10所阴阳结果判断完全按照试剂盒说明,而其他17所用酶标仪检测判断结果的实验室临界值则根据经验自行设置,吸光度(A)值、样本A值与临界值A值比值(S/CO值)均高于试剂盒推荐值.24所三级医疗机构中有10所受访者不了解抗HBs保护性浓度,另外14所的受访者知道抗HBs保护性浓度这个概念,但其中4所受访者知道得不准确,二级医院和私立机构对抗HBs保护性浓度则知之甚少.结论 抗HBs检测结果判读上存在着严重的主观性,抗HBs定性检测临界值的设置仍然没有统一,这可能是造成各实验室结果差异的主要原因,从业人员对专业知识的掌握有待加强.

Objective To compare the difference between ELISA and chemiluminescence assay in de-tecting hepatitis B surface antibody (HBsAb) serum valued in 5～100mIU/ml. Method 127 HBsAb serum samples that had valued by chemiluminescence were detected using 10 domestic ELISA kits. The correlation between the results of ELISA kits and chemiluminescence assay were analyzed. Coincidence between the results of ELISA kits and chemiluminescence at different OD value range were also analyzed. Results The detected results between 10 domestic ELISA kits and chemiluminescence assay are positive correlation (P<0.01); The optimal coincidence rates of the results of ELISA with those samples valued by chemiluminescence at more than 100mIU/ml was 99.4%; the coinci-dence rate is from 42% to 80% in cut-off±20%. The average coincidence rate is under 60%. Conclusions Though the detected results of 10 domestic ELISA reagents is related closely, the coincidence rate of the results at 5～10mIU/ ml in detecting anti-HBs is low. So it is important to make a positive standard in detecting anti-HBs.

目的 探讨使用浓度为10mIU/ml的乙肝表面抗体定值质控品作为乙肝表面抗体ELISA结果判断临界值的可行性.方法 采用6种国产主流HBsAb酶联免疫吸附试验(ELISA)试剂对86例经用化学发光法检测已知HBsAb浓度的标本进行检测,以浓度为10mIU/ml、30mIU/ml的乙肝表面抗体定值质控品和试剂盒标准为判断标准,分别比较不同浓度段和OD值段ELISA结果与化学发光法结果的符合率.结果 浓度分段比较显示:在5～10mIU/ml浓度段以质控品10mIU/ml和30mIU/ml的OD值为临界值符合率较高,分别足92.21%和91.07%.在10～30mIU/ml段以试剂盒判断标准、质控品10mIU/ml的OD值为临界值符合率较高,分别是80.49%和88.05%,在30～100mIU/ml段和≥100mIU/m段时,各判断标准的符合率大敛相同;以目前广州地区各实验室常用的临界值将OD值分段比较显示:在0.05～0.105段和0.105～0.21段,以10mIU/ml为临界值符合率较高,各试剂平均符合率达到74.39%,其余各段符合率大致相同;ELEISA OD值处于临界值±20%附近的符合率显示:以10mIU/ml的OD值为临界值平均符合率较为77.62%.以30mIU/ml的OD值为临界值时,平均符合率较为69.21%,CO-20%平均符合率比CO+20%平均符合率高.当以试剂盒标准为临界值时,平均符合率较为59.86%,CO+20%平均符合率比CO-20%平均符合率高.结论 使用10mIU/ml的定值质控品作为乙肝表面抗体ELISA结果判断依据得出的结果与化学发光结果符合率比使用试剂盒标准和30mIU/ml的定值质控品作为标准得出的结果高.浓度为10mIU/ml的乙肝表面抗体定值质控品应作为儿童乙肝表面抗体ELISA结果判断阴阳性的依据。

目的 探讨ANCA检测对川畸病(KD)的早期诊断、病情判断和预后转归等临床意义.方法 收集115例确诊川崎病患儿的实验室免疫学相关指标、彩色超声心动图检查和临床资料.采用SPSS11.0统计学软件比较ANCA阳性组与阴性组发病年龄、平均病程、冠状动脉损伤、复发情况及在丙球蛋白治疗前血清免疫球蛋白和补体浓度.结果 78例KD患儿血清中检测到ANCA,阳性率为67.8%.ANCA阳性与阴性组间在男士性别比例、疾病复发率上无明显差别,但是ANCA阳性组的平均发病年龄要早于阴性组,而平均病程则比阴性组要长;ANCA阴阳两组的补体C3、C4和免疫球蛋白IgA、IgM平均含量均处于正常参考范围内,且组间无差异,然而ANCA阳性组IgG平均含量(15.7±10.9g/L)明显高于阴性组.随着冠状动脉损伤程度的加重,ANCA阳性率也显著性提高.结论 ANCA为该病的一种常见自身抗体,有助于该病的早期诊断,同时对急性期的病程估计具有指导意义.ANCA与冠状动脉损伤程度的轻重密切相关,可能是通过激活补体乐统以外的途径参与冠状动脉病变的发生与发展.