Dried blood spots,valid screening for viral hepatitis and human immunodeficiency virus in real-life

AIM To detect chronic hepatitis B(CHB),chronic hepatitis C(CHC) and human immunodeficiency virus(HIV) infections in dried blood spot(DBS) and compare these samples to venous blood sampling in real-life.METHODS We included prospective patients with known viral infections from drug treatment centers,a prison and outpatient clinics and included blood donors as negative controls. Five drops of finger capillary blood were spotted on filter paper,and a venous blood sample was obtained. The samples were analyzed for HBs Ag,antiHBc,anti-HBs,anti-HCV,and anti-HIV levels as well as subjected to a combined nucleic acid test(NAT) for HBV DNA,HCV RNA and HIV RNA.RESULTS Samples from 404 subjects were screened(85 CHB,116 CHC,114 HIV and 99 blood donors). DBS had a sensitivity of > 96% and a specificity of > 98% for the detection of all three infections. NAT testing did not improve sensitivity,but correctly classified 95% of the anti-HCV-positive patients with chronic and past infections. Anti-HBc and anti-HBS showed low sensitivity in DBS(68% and 42%).CONCLUSION DBS sampling,combined with an automated analysis system,is a feasible screening method to diagnose chronic viral hepatitis and HIV infections outside of the health care system.

LC–HRMS determination of piperine on rat dried blood spots: A pharmacokinetic study

A liquid chromatography-high resolution mass spectrometry （LC-HRMS） method was developed and validated for the determination of piperine （PPR） on dried blood spots （DBS）. DBS samples were prepared by spiking the whole blood with analyte to produce 30 μL of blood spots on specimen collection cards. Chromatographic separation was achieved on an Atlantis dC18 column using acetonitrile and water （0.1% formic acid） （85：15, v/v） as mobile phase in an isocratic mode of elution at a flow rate of 0.75 mL/min. MS detection was carried out in electrospray positive ion mode for the target ions and monitored at m/z 286.1465 for PPR and 272.1303 for the internal standard （IS）. The developed method exhibited a linear dynamic range over 0.01-2000 ng/mL for PPR on DBS. The overall extraction recovery of PPR from DBS was 92.5%. Influence of hematocrit and spot volume on DBS was also evaluated and found to be well within the acceptable limits. The method was successfully applied to pharmacokinetic studies of PPR in rats.

Exploration of an Efficient Simultaneous Molecular Detection Method of HIV,HCV,and Syphilis from a Single Dried Blood Spot

Objective The aim of the present study was to evaluate the performance of the simultaneous detection of HIV-1 RNA,HIV-1 DNA,and HCV RNA using one dried blood spot(DBS)as an alternative sample to plasma.Method A total of 571 paired DBS/plasma samples were collected from men who have sex with men(MSM)and injection drug users(IDUs),and serological and molecular assays were performed.Using plasma results as the reference standard,the performance of DBS tests for HIV-1 RNA,HIV-1 DNA,and HCV RNA was evaluated.Pearson’s correlation coefficients and Bland-Altman analysis were performed to assess the correlation and concordance between DBS and plasma.Results Among paired plasma/DBS samples with detectable HIV-1 RNA and HCV RNA,five samples(5/32)were not detectable in DBS,while measurable HIV-1 RNA levels were present in plasma(1.44 to3.99 log10 copies/m L).There were two samples(2/94)with undetectable HCV RNA in DBS,while measurable HCV RNA levels were present in plasma(-5 to 5.99 log10 copies/m L).The correlation between HIV-1 RNA light chain variable region(VL)values obtained from plasma and DBS showed that r=0.683(P<0.01),n=27 and r=0.612(P<0.01),n=89 in HCV RNA.Bland-Altman analysis revealed that in HIV-1 RNA,the mean(±SD)difference between HIV-1 RNA in plasma and DBS was 1.00±1.01 log10 copies/m L,and all samples were within±1.96 SD(-0.97 to 2.97 log10 copies/m L)for DBS.The mean difference(±SD)in HCV RNA was 0.15±1.08 log10 copies/m L,and 94.38%(84/89)were within±1.96 SD(-1.96 to 2.67 log10 copies/m L).Overall,HIV-1 RNA and HCV RNA levels obtained from a DBS were lower than those obtained from plasma.HIV-1 DNA in a DBS showed concordant results with HIV-1 RNA in plasma.HIV-1 DNA RT-PCR using a DBS showed acceptable performance.Conclusion The performance of the simultaneous detection of HIV-1 RNA,HIV-1 DNA,and HCV RNA using one DBS was acceptable.DBS,as an alternative sample to plasma,may be a viable option for the simultaneous detection of HIV-1 RNA,HIV-1 DNA,and HCV RNA in resource-limited settings or for individuals living in areas that are difficult to access.

Dried blood spot sampling as an alternative for the improvement of hepatitis B and C diagnosis in key populations

BACKGROUND To achieve the elimination of hepatitis B and C,there is an urgent need to develop alternative strategies to increase the access of diagnosis,particularly among key populations such as people living with human immunodeficiency virus(HIV),individuals with coagulopathies and chronic kidney disease(CKD)patients.AIM To evaluate the use of dried blood spot(DBS)in the detection of hepatitis B virus(HBV)and hepatitis C virus(HCV)markers.METHODS A total of 430 individuals comprised of people living with HIV,coagulopathies and CKD provided paired serum and DBS samples.HBsAg,anti-HBc and anti-HCV were tested in those samples using a commercial electrochemiluminescence.Demographic and selected behavioral variables were evaluated to assess possible association with HBV and HCV positivity.RESULTS Using DBS,HBsAg prevalence varied from 3.9%to 22.1%,anti-HBc rates varied from 25.5%to 45.6%and anti-HCV positivity ranged from 15.9%to 41.2%in key populations.Specificities of HBV and HCV tests using DBS varied from 88.9%to 100%.The HBsAg assay demonstrated the best performance in CKD and coagulopathy individuals and the anti-HCV test had a sensitivity and specificity of 100%in people living with HIV.Accuracy of HBV and HCV detection in DBS varied from 90.2%to 100%.In the CKD group,HBsAg positivity was associated with infrequent use of condoms,and anti-HBc positivity was associated with sharing nail cutters/razors/toothbrushes.Anti-HCV reactivity was positively associated with a history of transplantation and length of time using hemodialysis in both specimens.In people living with HIV,only the male gender was associated with anti-HBc positivity in serum and DBS.CONCLUSION DBS with electrochemiluminescence are useful tools for the diagnosis and prevalence studies of hepatitis B and C among key populations and may increase the opportunity to foster prevention and treatment.

干斑法保存血浆中HBV、HCV、HIV病毒核酸及其保存效期研究

目的:建立一种以干斑法为基础制作的血液采集卡保存血浆中病毒核酸的方法,通过建立阿伦尼乌斯方程,预测该种保存方法保存核酸的效期,并论证该种保存方法在血站留存血液样本进行核酸复测的可行性。方法:分别将HBV、HCV、HIV核酸阳性的血浆样本制备成保存卡,以干浆斑的形式进行保存;将制备的保存卡分别放置于37、45、50和55℃条件下进行加速保存实验,不定期取出各个温度条件下的保存卡片,进行核酸提取操作,纯化出待测核酸样本,进行后续PCR测试,并记录CT值;根据阿伦尼乌斯方程建立起失效时间与储存温度之间的模型,进而推测出血液采集卡在规定保存温度条件下保存核酸的有效期。结果:HBV、HCV、HIV 3种核酸阳性的血浆样本,在使用干斑法保存时,其保存效期与温度的回归方程分别为:y=-11546x+31.74、y=-12949x+36.88、y=-12204x+34.48,线性回归的相关系数r均>0.98。预测HBV、HCV和HIV 3种病毒核酸使用干斑法保存时,在4℃的保存温度下,其保存效期分别为20792、19289和14285 d;在20℃的保存温度下,保存效期分别为2135、1502和1289 d。结论:血液样本中常见的3种病毒核酸,使用干斑法保存时,在加速破坏实验中符合一级反应模式,核酸降解速度与保存的绝对温度之间的关系符合阿伦尼乌斯方程。经过该方程的推算,使用干斑法保存血浆核酸样本,在不高于20℃的保存条件下,核酸稳定性效期最低可达3.5年,基本可满足血站血液样本留样保存年限的要求。

滤纸片干血斑在HIV-1 BED-CEIA新发感染检测方法中的应用

目的探讨在HIV-1 BED-CEIA新发感染检测中应用滤纸片干血斑的价值。方法研究纳入22个艾滋病自愿咨询检测(VCT)中心10226名咨询者血浆及干血斑样本作为目标实施HIV抗体检测,对通过免疫印迹法(WB)明确诊断为350例HIV感染患者的血浆样本及干血斑样本需同时实施BED-CEIA检测,观察滤纸片干血斑在HIV-1 BED-CEIA新发感染检测方法中稳定性、重复性,对两种样本检测结果存在的差异性进行对比。结果在HIV-1 BED-CEIA新发感染检测中应用滤纸片干血斑稳定性、重复性较高,重复性R^(2)值可高达0.9551。350例HIV感染患者样本检测结果中,其中295例患者样本被同时评估为长期感染,53例患者样本被同时评估为新近感染,两种样本对HIV BED-CEIA新发感染评估判定结果呈一致性(R^(2)值=0.95),一致性为99.42%。血浆样本及干血斑样本得到不同结果,样本An值均处于临界值附近(P<0.05)。结论在HIV-1 BED-CEIA新发感染检测中应用滤纸片干血斑的价值较高,检测结果稳定性、重复性较好,虽然部分样本检测结果存在一定的差异,但与血浆样本检测结果两者之间存在较佳的等效性,可广泛应用于临床HIV-1 BED-CEIA新近感染检测中。

串联质谱法和荧光分析法检测滤纸干血片苯丙氨酸的比较

目的比较荧光分析法和串联质谱法检测滤纸干血片上苯丙氨酸(Phe)水平的分布和差异,为实验室开展新生儿高苯丙氨酸血症(HPA)的筛查提供方法学依据。方法选择进行遗传代谢病筛查的滤纸干血片样本62 510例。采用荧光分析法和串联质谱法同步检测新生儿滤纸干血片的Phe浓度,同时还用串联质谱法检测酪氨酸(Tyr)浓度。采用配对Wilcoxon检验和等级相关分析评价2种方法间的差异和相关性,采用Bland-Altman一致性分析评价2种方法间的一致性。结果荧光分析法和串联质谱法均在62 510例新生儿滤纸干血片样本中检出3例HPA,敏感性均为100%。荧光分析法的阳性预期值为33.3%,串联质谱法的阳性预期值为18.8%,若联合Phe/Tyr比值,串联质谱法的阳性预期值可达100%。正常新生儿Phe浓度呈偏态分布,荧光分析法和串联质谱法检测结果<60μmol/L者分别占96.31%和95.08%,≥120μmol/L者仅占0.01%和0.03%。荧光分析法的测定值先低于串联质谱法,至97%位点(Phe≈63μmol/L)处基本持平,然后逐渐高于串联质谱法,2种方法的测定值呈正相关(r=0.43,P<0.01)。以不同浓度的Phe测定值作Bland-Altman分析,Phe浓度越高,2种方法的偏移越小,Phe>120μmol/L的19个样本点全部落在95%的一致性限内,2种方法有很好的一致性,检测结果有可替代性。结论荧光分析法和串联质谱法检测Phe在低浓度时有差异,高浓度时一致性好。2种方法对于HPA的临床判断无影响,均能用于新生儿筛查。串联质谱法可同时检测Phe和Tyr浓度,若联合Phe/Tyr比值,检出HPA的阳性预期值很高。

串联质谱检测干血滤纸片中氨基酸质控分析

目的通过对近3年的质控结果进行分析,探讨影响检测结果的因素及应对措施。方法美国疾病控制和预防中心(CDC)每年提供4批氨基酸质控干血滤纸片,利用串联质谱技术检测干血滤纸片中氨基酸水平。以美国CDC检测结果为标准,分析本实验室氨基酸质控结果偏差及其他实验室质控结果(213个实验室的均值)偏差之间的差异。结果本实验室每次质控结果所有氨基酸均在美国CDC检测结果95%可信区间内,且每一批结果的临床评价与美国CDC质控临床评价完全一致。本实验室质控结果偏差与其他实验室质控结果偏差比较,苯丙氨酸、亮氨酸、甲硫氨酸、酪氨酸和缬氨酸均无显著性差异,瓜氨酸有显著性差异(Z=-2.945P<0.005)。结论本实验室氨基酸质控结果,达到美国CDC的质控要求,瓜氨酸检测值偏低,其阳性判断切割值设定也相应降低。

串联质谱分析干血滤纸片中酰基肉碱质控

目的:串联质谱技术是近几年应用于临床检验的新技术,具有特异性强、灵敏度高,一次试验可检测数十种物质的优点。本文通过对近4年的质控结果进行分析,探讨影响检测结果的因素及应对措施。方法:美国CDC每年提供4批酰基肉碱质控干血滤纸片,酰基肉碱检测方法为取直径为3mm的质控干血滤纸片放入96孔聚丙烯板,经含酰基肉碱内标的甲醇萃取,盐酸正丁醇衍生后,进行串联质谱检测。分析本实验室质控结果与美国CDC检测结果的偏差及其他实验室质控结果(169个实验室的均值)与美国CDC检测结果的偏差之间的差异。结果:本实验室每次质控结果所有酰基肉碱均在美国CDC检测结果95%可信区间内,且每一批结果的临床评价与美国CDC质控临床评价完全一致。本实验室质控结果与美国CDC检测结果的偏差与其他实验室质控结果与美国CDC检测结果的偏差比较,无显著差异,丙酰基肉碱差异显著(Z=-2.638,P<0.005),戊二酰基肉碱差异亦显著(Z=-2.482,P<0.005)。结论:本实验室酰基肉碱质控结果,达到美国CDC的质控要求,丙酰基肉碱检测值偏高,其阳性判断切割值设定也相应增高,而戊二酰基肉碱检测值偏低,其阳性判断切割值设定也相应降低。

电喷雾串联质谱法检测干血斑中谷氨酰胺、谷氨酸及其临床初步应用

目的建立一种采用电喷雾串联质谱法(ESI-MS/MS)检测干血斑样品中谷氨酰胺(Gln)和谷氨酸(Glu)含量的方法,评价其用于筛查新生儿鸟氨酸氨甲酰转移酶缺乏症和氨甲酰磷酸合成酶缺乏症的可行性。方法收集2020年6—9月新生儿筛查足跟血干血斑样品,用非衍生化串联质谱检测试剂盒萃取样品,完成常规串联质谱检测项目后,将残余样品作为Gln和Glu的检测对象,复溶后选择正离子扫描模式进行分析测定,统计分析ESI-MS/MS检测Gln和Glu的正确度、精密度和稳定性,并评价其临床应用。结果Gln在125~1250μmol/L浓度范围内回收率和正确度均良好,回收率为98.01%~107.24%,偏倚为0.56%~4.8%;Glu在250~1250μmol/L浓度范围内回收率良好,回收率为102.06%~113.65%,在250~1250μmol/L浓度范围内正确度良好,偏倚为0.09%~7.67%。Gln精密度良好,Glu精密度稍差。稳定性实验证实在样品复溶前后,Gln差异无统计学意义,但Glu差异有统计学意义。结合Gln和Glu检测结果,6份鸟氨酸氨甲酰转移酶缺乏症确诊患儿干血斑样品检出率为100%,并确定Gln的参考区间为<106.63μmol/L,Glu的参考区间为<188.24μmol/L。结论成功建立了检测Gln和Glu的ESI-MS/MS法,该方法操作简单、成本低,不影响临床常规检测。

微量明胶颗粒凝集试验检测干血斑HIV-1抗体方法的建立与评价

目的建立微量明胶颗粒凝集试验检测滤纸片干血斑HIV-1抗体的方法,评价其敏感性和特异性。方法采集某吸毒哨点106份全血样本,制成滤纸片干血斑样本和血清样本;摸索微量明胶颗粒凝集试验中抗原明胶颗粒最佳稀释倍比;比较微量明胶颗粒凝集试验检测滤纸片干血斑和ELISA检测血清HIV-1抗体的结果。结果微量明胶颗粒凝集试验检测滤纸片干血斑HIV-1抗体的结果与ELISA检测血清HIV-1抗体的结果一致。结论微量明胶颗粒凝集试验检测滤纸片干血斑HIV-1抗体的方法,可信度较高、操作简便成本低,为大规模开展HIV-1流行病学调查提供了另一种良好的实验室方法。

新生儿干血斑β地中海贫血筛查方法的研究

目的探讨全自动毛细管微量血红蛋白电泳技术在新生儿β地中海贫血筛查中的应用价值,并寻求有效的筛查方法。方法使用Sebia全自动毛细管电泳仪对145904例新生儿滤纸干血斑标本进行血红蛋白(Hb)电泳;对95059例标本的电泳分析结果以HbA界值法判读β地贫筛查结果,对50845例标本的电泳分析结果以HbA值结合HbA2/HbA比值的综合分析法判读β地贫筛查结果,召回筛查阳性者进行地贫基因诊断;随机对600份标本的电泳结果同时以上述两种方法判读β地贫筛查结果,同时进行基因检测作为β地贫诊断的金标准,统计比较两种判读方法的差异。结果 HbA界值法筛查出的1580例阳性标本中经基因检测确诊为β地贫的有716例,确诊符合率45.32%,假阳性率为54.68%;综合分析法筛查出的1266例阳性标本中930例经基因检测确诊为β地贫,符合率73.46%,假阳性率为26.54%。同时对600例新生儿干血斑分析结果表明,HbA界值法和综合分析法的敏感度和特异度分别是54.55%、90.91%和97.58%、99.31%,两者的筛查准确性分别为96%和99%。统计结果表明,综合分析法的敏感度、阳性预测值和筛查的准确性显著高于HbA界值法.结论新生儿干血斑标本经毛细管血红蛋白电泳后,利用包含HbA、HbA2组分的含量及HbA2/HbA比值的综合分析法判读电泳结果,筛查新生儿β地贫,灵敏度和准确性显著提高,假阳性率明显降低,值得推广应用。

基质分散固相萃取-超高效液相色谱-串联高分辨质谱法同时测定干血斑样品中22种镇定催眠类药物

构建了干血斑样品中22种镇定催眠类药物的超高效液相色谱-串联高分辨质谱法定性、定量检测方法。干血斑样品经过乙腈提取后,应用基质分散固相萃取法对提取液进行净化,考察了不同种类净化剂以及用量对净化效果的影响,采用C18色谱柱进行分离,对色谱条件和质谱条件进行优化,使用同位素内标法进行定量分析。结果表明:22种被测物质在5 ng~800 ng/mL范围内线性表现良好(r^(2)>0.99),检出限为2 ng/mL~8 ng/mL,定量限为5 ng/mL~20 ng/mL,被测物质在低、中、高三个添加水平下回收率为80.2%~113.5%,相对标准偏差为0.9%~14.3%。本研究样本存储方便,保存时间长,前处理操作简单,应用精确质量数进行定性和定量,极大降低了假阳性的发生,为临床药物研究和法医毒物分析提供了新的技术支持。

Screening of amino acids in dried blood spots by stable isotope derivatization-liquid chromatography-electrospray ionization mass spectrometry

Direct infusion mass spectrometry(DIMS) is a powerful technique in clinical diagnosis for screening neonatal amino acid metabolic disorders from dried blood spots(DBS).However,DIMS sometimes generated false-positive results for analysis of amino acids.In this work,we utilized a stable isotope derivatization method,combining with liquid chromatography tandem mass spectrometry(SID-LC-MS),to improve the specificity for screening amino acids in DBS specimens.A pair of isotope reagents,p-(dimethylamino)phenyl isothiocyanate(DMAP-NCS) and 4-isothiocyanato-N,N-bis(methyl-[2H2])aniline([2H4]DMAP-NCS),was synthesized and used to label amino acids in DBS specimens.The [2H4]DMAP-NCS labelled amino acid standards were used as internal standards to compensate the matrix effect.This method was validated by measuring linearity,recovery and accuracy.The results showed that the developed SID-LC-MS method can be used for sensitive and selective determination of 12 diagnostically important amino acids in DBS specimens.

Detection of Schistosoma mansoni DNA using polymerase chain reaction from serum and dried blood spot card samples of an adult population in North-western Tanzania

Background:Real-time polymerase chain reaction(PCR)is a sensitive and specific method for diagnosing schistosomiasis.However,this method should be performed in a laboratory,usually located distant from the sample collection site.Therefore,it is important to have fast sampling preservation methods,which allow simple transport prior to DNA extraction and amplification.The aim of this study was to verify if blood samples applied to filter paper are suitable for analysis of Schistosoma mansoni DNA by real-time PCR.Methods:A cross-sectional study was conducted among 100 study participants aged 17 to 70 years in a fishing village on the southern shore of Lake Victoria,fanzania.Serum samples and ethylenediaminetetraacetic acid(EDTA)-anticoagulated whole blood for preparation of dried blood spots(DBS)were collected to test for Schistosoma mansoni infection by real-time PCR.A combined diagnostic reference of positive results of serum-based real-time PCR and the Kato-Katz(KK)method was used for analysis.Sensitivity and negative predictive value(NPV)were calculated.The Wilcoxon signed-rank test was chosen to compare the mean cycle threshold(Ct)values from serum and DBS.Results:According to the reference,92.5%S.mansoni positive samples were determined.The serum-based real-time PCR performed excellently with 95.4%sensitivity,whereas the DBS-based real-time PCR showed a low sensitivity(45.4%).The Ct-values were significantly higher in DBS(median:37.3)than in serum samples(median:27.5,P<0.001),reflecting a lower parasite-specific DNA load on the filter cards.With increasing egg counts,an increase in sensitivity was observed for all methods.The POC-CCA test and the serum-based real-time PCR showed a sensitivity of 100%for medium and severe infections.The DBS real-time PCR showed a sensitivity of only 85.7%even for severe infections.Conclusions:DBS-based real-time PCR did not provide good results in our study and therefore should not be recommended or must be tested concerning temperature of storage,storage duration,use of different filter papers and extraction methods before it is used in future studies.In contrast,our results showed that the POC-CCA test is a sensitive and precise test for detecting S.mansoni infections.

“新筛”干血斑在耳聋基因检测中的应用

目的将新生儿遗传代谢病筛查(简称“新筛”)后剩下的干血斑继续用于耳聋基因提取和飞行质谱检测,从而减少耳聋基因检测过程中的取样频次并缩短检测周期。方法在碱水煮沸法提取耳聋基因的基础上优化碱水的pH值和用量,使“新筛”干血斑DNA提取浓度达到新鲜血斑水平。结果用pH值为9、10、11、12的碱水从干血斑中提取DNA时,DNA的浓度与碱水pH值呈正相关(r=0.906,P=0.094),20个耳聋基因位点均能被正常检出;当pH值达到12时,质谱峰图信号响应值减小,检测质量下降。用60、70、80、90μL pH值为11的碱水从“新筛”干血斑中提取DNA时,DNA浓度与碱水体积呈负相关(r=-0.988,P=0.012);用90μL pH值为11的碱水时,有1例样本547G>A位点基因被漏检,用80μL pH值为11的碱水时,DNA提取浓度和新鲜血斑最接近(t=0.035,P=0.974),同时质谱峰图良好。结论用80μL pH值为11的煮沸碱水从“新筛”干血斑中提取DNA,可获得较好的耳聋质谱检测结果,为搭建“新筛”和耳聋自动化检测平台提供一定的技术参考。

在线干血斑结合2D-LC技术建立微量血浆中万古霉素测定方法

目的通过在线干血斑(DBS)结合2D-LC技术,建立自动化程度较高的人血浆中万古霉素测定方法,用于临床血药浓度监测工作。方法血浆样品离心后,取10μL滴在自制DBS滤纸上,冷风吹干5 min,制成干血斑试纸。将滤纸放在双层尼龙过滤膜中,用DBS适配器夹紧后连在2D-LC上自动进样。2D-LC中第一维(LC1)为反相色谱系统,第二维(LC2)为离子交换色谱系统,LC1及LC2系统通过六通阀连接,采用"中心切割"模式及陷阱柱转移目标物,检测波长为280 nm。结果血浆样品制成DBS后稳定性较高,便于长时间保存和长途运输。所建立的2D-LC方法具有很强的抗干扰能力,运行受环境影响因素极小,因此无需内标校正测定误差。万古霉素在0.5~60μg·mL^-1与峰面积线性关系良好,相关系数为0.9999,LOQ为0.200μg·mL^-1。萃取回收率在93.6%~95.3%,日内及日间精密度RSD均<6%,准确度在98.3%~99.7%。结论 DBS能有效解决血浆样品长距离与长时间稳定性问题,测定速度快,血样需要量小,适用于新生儿或长距离样品运输的万古霉素血浆药物浓度检测。

分析筛查新生儿干血斑β地中海贫血的方法及效果

目的:探讨全自动毛细管微量血红蛋白电泳技术在新生儿β地中海贫血筛查中的应用价值。方法:收治新生儿145 902例,采用Sebia全自动毛细管电泳仪进行血红蛋白电泳。对95 058例标本以Hb A界值法读出β地中海贫血筛查结果,对50 844例标本根据Hb A值结合Hb A2/Hb A比值读出β地中海贫血筛选结果。随机选取598份结果判读β地中海贫血筛选结果,同时基因检测为β地中海贫血金标准。结果:Hb A界值法β地中海贫血符合率45.2%、假阳性率54.8%;综合分析法β地中海贫血符合率73.5%,假阳性率26.5%。Hb A与综合分析敏感度与特异度分别54.54%、90.90%和97.57%、99.30%。综合分析法在阳性预测值与筛选准确性等方面高于Hb A界值法。结论:综合分析法在筛选新生儿β地中海贫血,判断电泳结果上灵敏度与精确度等较高。

Comparative study on anti-HCV testing using plasma, dried plasma spots (DPS), and dried blood spots (DBS)

The aim of this study was to evaluate the performance of an assay using dried plasma spot(DPS)and dried blood spot(DBS)samples for the serological detection of anti-hepatitis C virus(HCV)antibodies.Between January and July 2019,plasma,DPS and DBS specimens were collected from individuals at high-risk for HCV infection.Samples were tested for anti-HCV by ELISA,and the performance of DPS and DBS specimens was examined using results from the plasma testing,as the standard.Blood samples were collected from 329 persons,including 129 men who have sex with men and 200 intravenous drug users.Results from the plasma testing indicated that 118 samples(59.0%)were HCV positive.Data from the DPS sample testing showed sensitivity as 99.2%(95%confidence interval[CI]:0.95-1.00)and specificity as 100%(95%CI:0.98-1.00)for HCV detection,with Kappa of 99.3%(95%CI:0.98-1.00)while in DBS sample testing the sensitivity as 98.3%(95%CI:0.93-1.00)and specificity as 100%(95%CI:0.98-1.00),with Kappa of 98.7%(95%CI:0.97-1.00),respectively.Spearman’s correlation coefficients for the comparisons between plasma and DPS specimen,plasma and DBS specimens,DPS and DBS specimens were 0.857,0.750,and 0.739,respectively.Compared with the results in plasma,1 sample was not detected using the DPS specimens,and 2 samples were failed for the positive detection,using the DBS specimens.Both DPS and DBS samples were promising alternatives to plasma,for the detection of anti-HCV antibodies.

摩梭人身体铁状态与血糖升高的相关性

目的探讨中国云南摩梭人群生理性铁储备,贫血与血糖升高的相关性。方法收集云南省宁蒗县187名摩梭人血样,检测其糖化血红蛋白(HbA1c)、转铁蛋白受体(TfR)与血红蛋白水平(Hb)。逻辑回归分析摩梭人数据和中国全国性数据(CHNS)中铁状态与血糖升高的关系。结果与控制组相比,摩梭人缺铁而不贫血,即缺铁性红细胞生成(IDE)对应着血糖升高风险的上升(OR=2.70),这一结果与针对全国性数据的分析结果一致。结论相比健康人群,缺铁性贫血与非缺铁性贫血的摩梭人,缺铁而不存在贫血症状的摩梭人面临着血糖升高的风险。摩梭人身体铁水平对其2型糖尿病的风险评估与预防具有参考意义。