UF-50联合干化学法在尿RBC、WBC检测中的应用价值

目的：探讨UF-50、尿干化学法与显微直接镜检法在尿RBC、WBC检测中的应用价值。方法：对尿RBC、WBC检测采用UF-50、尿干化学法与显微直接镜检法。结果：三种检测方法结果为高度相关，RBC符合率89.2%，K值0.846；WBC符合率为90.9%，K值0.858。RBC的检出率为UF-50〉干化学法〉镜检法；WBC的检出率为UF-50〉镜检法〉干化学法。结论：UF-50对尿沉渣RBC、WBC有较高灵敏度，三种检测方法互补，联合应用可大大降低人工镜检复检率，但对UF-50与干化学法结果不符或可疑的标本，一定要通过标准化显微镜复检。     

全自动尿沉渣仪分析法与尿沉渣显微镜镜检的应用价值

[目的]探讨尿沉渣分析仪、尿干化学分析仪及涂片显微镜检测方法的临床应用价值。[方法]采用UF-100型全自动尿沉渣分析仪、JUNNUR—Ⅱ尿干化学分析仪、手工涂片尿沉渣显微镜镜检对560份住院患者的尿液标本沉渣进行检测。[结果]尿干化学法、尿沉渣分析仪法及显微镜镜检法检测白细胞阳性率分别为55．18％、58．57％、56．25％,检测红细胞阳性率分别为63．21％、60．18％、56．43％,差异无统计学意义（P〉0．05）。[结论]尿干化学法、尿沉渣分析仪法及显微镜镜检法检测具有很高的准确性和敏感度,为泌尿生殖系统、循环系统、内分泌系统等疾病的诊断提供了可靠的数据。     

SM染色法等在泌尿系感染中的诊断价值

目的探讨SM尿沉渣染色镜检在泌尿系感染疾病中的诊断价值。方法第一组1834例样本用UF-100尿沉渣分析仪、干化学分析仪、SM染色镜检三种方法对尿中白细胞进行分析,第二组2730例样本镜检用传统的非染色法。结果第一组UF-100尿沉渣仪法、干化学法、SM染色镜检法检测WBC阳性率分别为32.6%、19.1%、23.6%。第二组UF-100尿沉渣仪法、干化学法、传统的非染色镜检法检测WBC阳性率分别为31.5%、18.2%、15.5%。结论UF-100尿沉渣分析的WBC检出率较高,但易受结晶、小圆上皮细胞因素等影响;干化学分析WBC检出率较低,干扰因素较多,且WBC数较高范围时,干化学分析法不能区分其变化;非染色法镜检检出率低;SM染色后镜检能更好地反映WBC总数和活WBC数的变化。UF-100尿沉渣仪与干化学法检测尿WBC结果不一致时,一定要进行尿沉渣染色镜检,才能提高尿沉渣检验质量,为临床诊断急、慢性泌尿系感染和疗效判断提供指导。     

SM染色法等在泌尿系感染中的诊断价值

目的探讨SM尿沉渣染色镜检在泌尿系感染疾病中的诊断价值。方法第一组1834例样本用UF-100尿沉渣分析仪、干化学分析仪、SM染色镜检三种方法对尿中白细胞进行分析,第二组2730例样本镜检用传统的非染色法。结果第一组UF-100尿沉渣仪法、干化学法、SM染色镜检法检测WBC阳性率分别为32.6%、19.1%、23.6%。第二组UF-100尿沉渣仪法、干化学法、传统的非染色镜检法检测WBC阳性率分别为31.5%、18.2%、15.5%。结论UF-100尿沉渣分析的WBC检出率较高,但易受结晶、小圆上皮细胞因素等影响;干化学分析WBC检出率较低,干扰因素较多,且WBC数较高范围时,干化学分析法不能区分其变化;非染色法镜检检出率低;SM染色后镜检能更好地反映WBC总数和活WBC数的变化。UF-100尿沉渣仪与干化学法检测尿WBC结果不一致时,一定要进行尿沉渣染色镜检,才能提高尿沉渣检验质量,为临床诊断急、慢性泌尿系感染和疗效判断提供指导。     

尿干化学法与沉渣镜检联合检测尿红细胞及白细胞的临床意义

目的分析尿干化学法与沉渣镜检的临床应用价值,探讨为患者带来更加准确、及时的检测结果途径。方法采用南京千盛医疗设备有限公司QS8005型流式尿沉渣分析仪(尿沉渣镜检),华晟H-Ⅱ尿液分析仪(尿干化学法)对300例尿液标本进行检测。结果尿干化学法检测白细胞假阳性率1.99%,假阴性率16.33%;红细胞假阳性率8.78%,假阴性率5.26%。两种方法对尿液白细胞阳性率进行检测,差异无统计学意义(P>0.05);对红细胞阳性率进行检测,差异有统计学意义(P<0.01)。结论尿干化学法与沉渣镜检白细胞和红细胞具有一定的互补性,两者联合检测能大大提高工作效率和检测结果准确度。     

不同方法检测尿红细胞的对比分析

目的探讨不同方法检测尿红细胞时的差异性,比较不同仪器检测方法对红细胞尿的携带污染率情况。方法对来自临床1 000例尿液标本分别用URISYS2400干化学分析仪、UF-500i尿有形成分分析仪、显微镜镜检进行检测。对健康成人尿标本和与之混合的不同浓度红细胞悬液交叉排列分别用干化学法和尿沉渣法进行检测。结果分别用干化学法、尿沉渣法、显微镜镜检检测尿红细胞的阳性率为41.1%(411/1 000)、22.6%(226/1 000)、19.7%(197/1 000),干化学法和尿沉渣法分别与显微镜镜检比较复合率分别是76.8%(768/1 000)、97.1%(971/1 000)。干化学和尿沉渣同时红细胞检测阴性为512例,采用显微镜复检均为阴性。携带污染率URI-SYS2400干化学分析仪为0.000%、UF-500i尿有形成分析仪为0.113%。结论干化学法、尿沉渣法检查尿红细胞可以提高检测效率,两者相互补充可以提高检测的准确度,但仍有影响因素不可避免,所以当两者检测不符时,仍然需要显微镜镜检确认。     

干化学法联合沉渣镜检检测尿液红细胞和白细胞的比对研究

目的探讨干化学法对尿液中红细胞(RBC)和白细胞(WBC)检测的筛查意义。方法通过干化学法和沉渣镜检对600份新鲜尿液标本进行检测,对其检测尿液RBC和WBC结果进行比对分析。结果干化学法尿液RBC阳性228例,阳性率为38.0%,沉渣镜检尿液RBC阳性194例,阳性率为32.3%,两者差异有统计学意义(χ2=4.225,P=0.040);干化学法尿液WBC阳性63例,阳性率为10.5%,沉渣镜检尿液WBC阳性93例,阳性率为15.5%,两者差异有统计学意义(χ2=6.631,P=0.010)。结论干化学法检测RBC和WBC时,有较高的假阳性率和假阴性率,干化学法只能作为尿液常规检查的筛查作用,不能完全替代沉渣镜检法。     

尿常规检查在泌尿系结石患者中的应用

目的了解红细胞、草酸钙结晶在泌尿系结石患者尿中出现的情况。方法采集经B超诊断的176例泌尿系结石患者新鲜尿液，用干化学法检测红细胞，尿离心镜检红细胞和草酸钙结晶。结果干化学法红细胞阳性率92％，尿离心镜检红细胞阳性率82％，草酸钙结晶阳性率74％。结论尿常规检查主要是检查尿红细胞，但草酸钙结晶在泌尿系结石患者中也有较高的阳性检出率。     

儿童尿液分析与尿沉渣联合检测的应用价值

目的探讨儿童尿液分析与尿沉渣联合检测的应用价值。方法采用日本东亚Sysmex UF-100全自动尿流式分析仪和韩国盈东Uriscan-Pro尿液干化学分析仪和沉渣镜检3种方法分析1000例尿液标本。结果UF-100尿流式分析仪与镜检符合率较高。结论UF-100可弥补干化学法检测的不足，显微镜可排除UF-100的假阳性及假阴性。     

干化学法和显微镜法检测尿液的对比分析

目的：探讨干化学法与显微镜镜检法检测尿液中红细胞、白细胞结果差异。方法：采集1 458例住院患者新鲜晨尿15 mL,采用干化学法和显微镜镜检法进行检测。结果：尿干化学法检测红细胞,1 186份阴性结果中镜检阳性为26份,占2.2%,272份阳性结果中镜检阴性33份,占12.1%;尿干化学法检测白细胞,817份阴性结果中镜检阳性为58份,占7.1%641份阳性结果中镜检阴性60份,占9.4%。结论：干化学法与显微镜镜检法检测尿液中红细胞、白细胞各有优缺点,只有将二者很好地结合使用,才能为临床提供正确的检验报告。     

外周静脉抗凝血沉淀试验在羊水栓塞早期确诊中的价值

目的 探讨外周静脉抗凝血沉淀试验在羊水栓塞早期确诊中的价值,为复苏与救治提供依据.方法 采用血液沉淀试验,通过回顾分析,对2005年3月～2014年3月发生在蒲城县医院和其他医院的10例羊水栓塞病人（观测组）的静脉抗凝血,离心后取血浆上层及全血底层样本,分别做羊水成分检测,并对同期该院随机抽取的100例健康产妇（对照组）破膜后至产后2h内的抗凝血标本做同样检测.结果 观测组10例,6例（60％）血浆上层和全血底层均检测到脂肪球、毳毛、鳞状上皮细胞、胎粪、黏液和碎片,3例（30％）检出脱落上皮和鳞状上皮,1例（10％）检出脂肪球和毳毛,阳性检出率100％.对照组100例,仅1例（1％）标本有极少量脂肪球,其余均未见任何羊水成分,阳性检出率1％.经统计学处理,两组检出率差异有统计学极显著性意义（X2=192,P＜0.005）.结论 外周静脉抗凝血离心沉淀试验对羊水成分的检出,快速、简单、经济.具有更早期确诊和警示栓塞发生的价值,可在临床中推广.     

羊水量异常的超声诊断与胎儿畸形的关系

目的 探讨羊水量异常与胎儿畸形的关系.方法 回顾分析2004年3月至2006年11月在本院进行二维及三维超声检查34 900例20周以上的胎儿,其检出羊水量异常160例,并研究分析羊水量多少与胎儿畸形的关系.结果 160例羊水量异常中,羊水过多100例,羊水过少60例,二者畸形的发生率分别为37%、31.6%.结论 超声是诊断羊水量异常的首选方法,而且还能发现胎儿畸形,进一步证明羊水量异常与胎儿畸形的发生密切相关.     

多重探针连接依赖式扩增快速检测染色体非整倍体异常

目的评价多重探针连接依赖式扩增（multiplex ligation-dependent probe amplification，MLPA）在常见染色体非整倍体异常及其在产前诊断中的应用价值。方法收集经染色体核型分析证实的12例21三体、1例（45，X）、1例（47，XYY）患者，8名正常健康人外周血标本和4例21三体胎儿脐带血标本及1例21三体胎儿羊水、7例核型正常胎儿羊水，共计34份样本。提取外周血或脐带血基因组DNA，羊水标本蛋白酶K消化获得细胞裂解液，采用MLPA技术检测34份标本13、18、21、X和Y染色体拷贝数的变化。结果48h内即可完成样本分析。除1份离心后含大量红细胞的羊水标本经MLPA分析后提示（69，XXY）或母血污染外，其余33份外周血、脐带血或羊水标本报告均与染色体核型分析结果一致，临床符合率97．1％。正常二倍体标本Ratio值除Y染色体相对拷贝数变异略大外，其余均接近于1．0。经单因素方差分析，21三体和正常二倍体标本的Ratio均值间差异有统计学意义（F=298．906，P=0．000）。结论高灵敏度MLPA技术结合计算机辅助数据分析方法操作简便、快速、可自动化，是诊断常见染色体非整倍体异常的可靠方法。     

Testing for maternal cell contamination in prenatal samples: a comprehensive survey of current diagnostic practices in 35 molecular diagnostic laboratories

The potential presence of maternal cell contamination (MCC) in chorionic villus or amniotic fluid samples poses a serious preanalytical risk for prenatal misdiagnosis. The aim of this study was to identify current diagnostic practices in the absence of comprehensive practice guidelines. Thirty-five clinical molecular laboratories that conduct prenatal testing agreed to participate in a clinical practice survey. The survey included questions about sample requirements, test indications, assay type, test performance and limitations, criteria and management of uninformative test results, reporting, and billing. Sixty percent of participating laboratories performed testing on direct and cultured amniotic fluid, whereas forty percent tested cultured cells only. Most also accepted chorionic villus samples. Although MCC testing of fetal samples is recommended in guidelines by the American College of Medical Genetics, only 60% of surveyed laboratories performed it without exception. Commercially available assays were used by 75% of participating laboratories, and at least five identity markers were evaluated at 87% of the laboratories. The reported lower limit of MCC detection ranged from 1 to 20% but was not determined in all laboratories. MCC testing was performed in the majority of molecular diagnostic laboratories, but guidelines for standardization are needed to ensure optimal and accurate prenatal patient care.     

HBV宫内感染机制的研究

目的：研究HBV宫内感染机理。方法：应用PCR技术检测59例HBsAg( )孕妇血清、阴道分泌物、羊水、胎盘及新生儿脐血清中HBV DNA。采用免疫组化ABC染色法检测胎盘中HBsAg及HBeAg。结果：新生儿宫内感染率为45．8％(27／59)；胎盘、羊水、阴道分泌物HBV DNA( )者宫内感染率分别为78．8％(26／33)、78．6％(22／28)、80．6％(25／31)均显著高于其HBVDNA(-)者。免疫组化ABC染色法检测胎盘中HBsAg、HBcAg。HBsAg及HBeAg在胎盘中分布由蜕膜细胞、滋养层细胞、绒毛间质细胞至胎儿面绒毛毛细血管内皮细胞依次递减，有胎盘中HBsAg及HBeAg分布于上述方向相反。当羊水、阴道分泌物HBV DNA( )时羊膜上皮细胞中检出HBsAg及HBcAg。结论：HBV经母血和(或)细胞直接蔓延方式进入胎儿血循环感染胎儿；可能存在阴道上行感染。     

Typing of Fetal Cells Obtained by Amniocentesis

In 38 of 287 amniocenteses done at the Mayo Clinic the amniotic fluid contained enough erythrocytes to permit blood typing with commercial sera. Simultaneous use of both an acid-elution technic and the direct Coombs test helped to establish three of the four possibilities for having erythrocytes in bloody amniotic fluid: 1) maternal and fetal erythrocytes of different blood types; 2) maternal erythrocytes only; 3) fetal erythrocytes only; and 4) maternal and fetal erythrocytes of the same blood type. Twenty of the 38 bloody amniotic fluids permitted fetal erythrocyte-typing that was later verified postnatally in the living newborns. Traumatic amniocentesis in an Rh_o(D)-negative mother with an Rh_o(D)-positive fetus increases the risk of anamnestic rise in maternal antibody levels. Amniotic epithelial cells from seven different nonbloody amniotic fluids were typed by mixed agglutination technic. Although the ABO groupings were accurate as shown by postnatal typing, Rh_o(D) typing was unreliable.     

Evaluation and quality control of a monoclonal antibody based enzyme antigen immunoassay of acetylcholinesterase in amniotic fluid

We have evaluated a new monoclonal antibody based enzyme antigen immunoassay (EAIA) for acetylcholinesterase in amniotic fluid, intended for the detection of open fetal abnormalities, especially open neural tube defects. With a sample volume of 50 microliters, the detection limit is 0.7 nkat/l and linearity is found up to an amount 200-times the detection limit. CV's are less than or equal to 6.6% within assays and less than or equal to 10.3% between assays; recoveries averaged 103%. The upper limit of the reference range for amniotic fluids from non-affected pregnancies is 8.5 nkat/l for clear specimens and 25.0 nkat/l for haemolysed specimens. Amniotic fluid specimens from 1473 patients (1388 normal and 85 pathological pregnancies) were examined with both the new EAIA and the original procedure using a polyclonal rabbit antiserum. The two procedures showed identical sensitivities and specificities of about 100%, except for amniotic fluid samples containing haemolysed blood. For the latter the new EAIA showed a specificity of 100% compared to 55.6% for the original procedure. We conclude that the new EAIA is accurate and reproducible and shows, compared with the original procedure, an increased specificity in the analysis of amniotic fluid samples containing haemolysed blood.     

BC-6900全自动血液细胞分析仪检测外周血有核红细胞的准确性分析

目的探讨BC-6900全自动血液细胞分析仪法(简称仪器法)检测外周血有核红细胞(NRBC)的方法学特点,评价其临床应用价值。方法同时用仪器法和显微镜检查法(简称镜检法)检测100例外周血标本中的NRBC,以镜检法为金标准,比较不同方法的计数结果,并从批内精密度、稳定性、携带污染率、人机比对方面分析BC-6900全自动血液细胞分析仪检测NRBC的效果。结果仪器法检测NRBC的灵敏度为97.959%(48/49),特异度为96.078%(49/51),假阳性率为3.922%(2/51),假阴性率为2.041%(1/49)。仪器法与镜检法检测结果有极好的相关性(r=0.9731),2种检测法比较,NRBC计数结果差异无统计学意义(P>0.05)。仪器法检测NRBC的重复性好,线性范围较宽,携带污染率小,比对试验中各项目变异系数(CV)均小于实验室允许的CV。结论BC-6900全自动血液细胞分析仪检测外周血NRBC性能良好,能满足临床医生对全血检测分析的需求。     

Microsatellite markers within --SEA breakpoints for prenatal diagnosis of HbBarts hydrops fetalis

BACKGROUND: We sought to develop a rapid prenatal diagnostic test for simultaneous detection of HbBarts hydrops fetalis and exclusion of maternal contamination. METHODS: We developed a multiplex quantitative fluorescent PCR (QF-PCR) test that detects the presence/ absence of 2 microsatellite markers (16PTEL05/16PTEL06) located within breakpoints of the Southeast Asia ((-SEA)) deletion. HbBarts hydrops fetalis ((-SEA/-SEA)) is diagnosed by absence of both markers, and maternal contamination of fetal DNA is excluded by absence of noninherited maternal alleles. Fetal and parental DNA samples from 50 families were analyzed in a blinded clinical validation study, and QF-PCR results were compared with their respective molecular genotypes. RESULTS: The multiplex QF-PCR results included correct diagnoses of HbBarts hydrops fetalis in 11 of the fetuses tested, correct verification as unaffected in 20 fetuses, and correct identification as either carriers (alphaalpha/(-SEA)) or unaffected homozygotes in 18. Misidentification as unaffected occurred for 1 carrier. Sensitivity for diagnosis of HbBarts hydrops fetalis was 100% [lower 95% confidence interval, 76.2%], and specificity was 100% (lower 95% confidence interval, 92.6%). None of the samples tested showed any traces of noninherited maternal alleles; thus false-positives because of maternal contamination were eliminated. CONCLUSIONS: In this QF-PCR method, detection of maternally and paternally inherited fetal alleles allowed diagnosis of the double-deletion syndrome, and the ability to differentiate between these alleles allowed simultaneous exclusion of maternal contamination of the fetal genetic material. This novel strategy using cell-free fetal DNA in maternal plasma could form the basis for noninvasive testing for HbBarts hydrops fetalis.     

孕妇外周血胎儿细胞中巨细胞病毒基因的检测及意义

目的利用孕妇外周血中胎儿细胞进行无创性产前诊断巨细胞病毒宫内感染的新方法。方法(1)采用显微操作技术从273例孕妇外血中分离单个胎儿有核红细胞。(2)应用多重引物原位合成技术(primed in situ labeling，PRINS)检测76例孕妇外周血HCMV-DNA阳性标本的单个胎儿细胞的SRY基因和HCMV-DNA基因。(3)应用引物延伸预扩增法(primed extension preamplification，PEP)及聚合酶链反应(PCR)检测273例孕妇外周血中单个胎儿细胞的SRY基因和HCMV-DNA基因。结果(1)应用显微操作技术分选胎儿细胞的分选率为100％。(2)PRINS技术检测SRY基因的敏感性为97．56％(40／41)，特异性为100％(35／35)。检测HCMV-DNA基因的敏感性为92．68％(38／41)，特异性为100％(35／35)。(3)PEP及PCR方法检测SRY基因的敏感性为97．39％(149／153)，特异性为99．17％(119／120)，检测HCMV-DNA基因的敏感性为95．12％(39／41)，特异性为100％(232／232)。结论应用PRINS技术、PEP方法及PCR技术对孕妇外周血中单个胎儿细胞进行非创伤性产前诊断巨细胞病毒宫内感染是一种敏感性高特异性强的新方法，可能具有广阔的临床应用前景。