Active tracking of rejected dried blood samples in a large program in Nigeria

AIM: To study the impact of rejection at different levels of health care by retrospectively reviewing records of dried blood spot samples received at the molecular laboratory for human immunodeficiency virus(HIV) early infant diagnosis(EID) between January 2008 and December 2012.METHODS: The specimen rejection rate, reasons for rejection and the impact of rejection at different levels of health care was examined. The extracted data were cleaned and checked for consistency and then deduplicated using the unique patient and clinic identifiers. The cleaned data were ciphered and exported to SPSS version 19(SPSS 2010 IBM Corp, New York, United States) for statistical analyses.RESULTS: Sample rejection rate of 2.4%(n = 786/32552) and repeat rate of 8.8%(n = 69/786) were established. The mean age of infants presenting for first HIV molecular test among accepted valid samples was 17.83 wk(95%CI: 17.65-18.01) vs 20.30 wk(95%CI: 16.53-24.06) for repeated samples. HIV infection rate was 9.8% vs 15.9% for accepted and repeated samples. Compared to tertiary healthcare clinics, secondary and primary clinics had twofold and three-fold higher likelihood of sample rejection, respectively(P < 0.05). We observed a significant increase in sample rejection rate with increasing number of EID clinics(r = 0.893, P = 0.041). The major reasons for rejection were improper sample collection(26.3%), improper labeling(16.4%) and insufficient blood(14.8%). CONCLUSION: Programs should monitor pre-analytical variables and incorporate continuous quality improvement interventions to reduce errors associated with sample rejection and improve patient retention.

Ethical and Regulatory Issues with Residual Newborn Screening Dried Bloodspots

After newborn screening is completed, most states retain leftover dried bloodspots. These dried bloodspots are stored for varying lengths of time among different state newborn screening programs. Dried bloodspots are a unique and valuable resource for the development of new newborn screening tests, quality assurance and biomedical research. Recent changes to the 2014 Newborn Screening Reauthorization Saves Lives Act require explicit parental consent for the retention and use of dried bloodspots in federally funded research. This has raised several ethical and regulatory issues and highlighted the challenges of respecting individual autonomy and public health goals. This article provides an overview of these issues and discusses methods for obtaining parental consent. These issues may be applicable to consent for the storage and use of biospecimens among other settings according to proposed changes to the Common Rule.

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.

适用于法庭科学毒物分析的干血斑检验体系的建立——以5种常见药(毒)物为例

干血斑技术能够方便地对血液样品中的违禁药物进行快速分析,在酒后驾驶检查、滥用药物检测、兴奋剂检测等毒物分析场景具有显著优势。然而在我国法庭科学毒物分析领域,因缺少标准化检验体系,其稳定性和可靠性未得到深入研究论证,限制了其在司法实践中的运用。本研究以甲基苯丙胺、利多卡因、氯胺酮、芬太尼和地西泮为典型药(毒)物,使用整个干血斑进行分析,建立了适用于法庭科学领域毒物分析的超高效液相色谱-串联质谱分析方法,形成了以干血斑样品制作、前处理、分析、储存和效用性评价为主要内容的检验体系,并为干血斑中其他药(毒)物的分析方法开发提供参考。结果表明,干血斑中利多卡因和芬太尼在0.5~100 ng/mL内线性关系良好,甲基苯丙胺、氯胺酮、地西泮在2~100 ng/mL内线性关系良好,方法检出限为0.2~0.5 ng/mL。干血斑中5种目标物可以在60天内保持稳定,目标物测定含量与理论值的偏差在15%以内。干血斑中5种目标物的测量结果与全血一致,没有显著的系统误差和比例误差,芬太尼、地西泮、氯胺酮、利多卡因和甲基苯丙胺的测量浓度的相对偏差分别为4.44%、3.50%、7.66%、5.10%和5.25%。干血斑样品前处理方法简单,样品用量小,能够实现血液样品保存的轻量化和规范化且与全血样品具有高度定量一致性,可为公安实践工作中分析、保存血液检材提供新方案。

干血片用于HCV抗体及RNA检测的评价

目的研究干血片在用于检测HCV抗体及RNA的性能评价。方法采集191例血标本制成血浆和干血片样本,用酶联免疫吸附试验(ELISA)检测HCV抗体以及用RT-PCR检测HCV-RNA;并将血浆检测HCV抗体和HCV-RNA阳性标本的干血片分别在不同温度条件下保存不同时间检测。结果干血片检测HCV抗体和HCV-RNA的灵敏度分别为97.8%、94.9%,特异度分别为100%、99.1%,与血浆比有很好的一致性;干血片在-20℃或4℃可以保存1年以上、25℃保存3个月HCV抗体结果稳定;干血片-20℃或4℃保存6个月、25℃保存1个月HCV-RNA结果稳定。结论用干血片样本检测HCV抗体及RNA与血浆样本检测结果差异无统计学意义,是血浆检测的可靠替代品,而且它在低温下保存时间长、常温也较稳定,方便保存、运送,比血浆适用范围更广范。

临床干血斑样本中12种抗凝血鼠药的液相色谱－串联质谱法同时定量检测

为应对临床中毒快检的需求,建立了干血斑样本中12种抗凝血鼠药的液相色谱-串联质谱定性定量方法。针对全打孔的干血斑样本,考察了滤纸卡种类、润湿步骤、提取溶剂种类对提取效果的影响。采用C18色谱柱进行分离,以乙酸铵(5 mmol/L)水溶液-乙酸铵(5 mmol/L)甲醇溶液为流动相进行梯度洗脱,在电喷雾负离子电离方式下使用三重四极多反应监测模式检测。结果表明,采用Whatman903滤纸卡为基底,以水为溶剂充分润湿,再以含内标的甲醇溶液提取5 min,各鼠药的提取率为66%~115%,且结果稳定(日内RSD小于15%)。除杀鼠酮的线性范围为20~500 ng/m L(r2=0.998 7)外,其它11种鼠药的线性范围为5~500 ng/m L(r2=0.998 8~0.999 6);12种鼠药的回收率为61%~105%,基质效应为71%~193%(日内RSD小于15%)。该方法准确、灵敏、快速且操作简便,成功应用于3例临床鼠药中毒病人样本的快速检测,为临床毒物检测和法医毒物分析的快速筛查和准确定量提供了新的技术方法。

干血点采样设备及方法在兴奋剂检查中的应用

目的:通过对运动员指尖或上臂开展干血点(driedbloodspot,DBS)器材测试,比较不同采血设备和采血针头搭配效果,探讨符合世界反兴奋剂机构(WADA)样本采集要求的DBS采样方式,为DBS技术在兴奋剂检查领域的推广应用做准备。方法:选取12名检查官对60名覆盖奥运会和冬奥会大部分兴奋剂高危项目的运动员进行不同组合方式的DBS采样,选取Tasso、Hemaxis、HemaSpot、Capitainer及金豪5种采血设备,搭配施莱低血量(SL)、施莱中血量(SM)、施莱高血量(SH)、BD低血量(BDL)、BD中血量(BDM)、BD高血量(BDH)6种采血针头,对运动员在采集过程中对兴奋剂检查方式偏好、采样痛感、出血情况、采血时间、失败情况等数据进行统计分析。结果:1)相比传统兴奋剂尿检、血检检查,运动员更倾向于DBS采样方式(P<0.01);2)BDL针头采血合格率最低,与其他5种型号针头相比均具有显著性差异(P<0.01),BDL针头采血量无法满足WADA定量采血要求;手臂血采集合格率低于指尖血合格率,BDM、BDH采血针采血合格率高于施莱3种采血针,但不存在差异(P>0.05);3)手臂血采血时间显著高于指尖血采血时间(P<0.01),各指尖采血方式用时无差异;4)满足采血量要求的情况下,BDH针头按摩手指次数明显少于其他型号针头(P<0.01),BDM针头按摩次数少于SM针头(P<0.01)和BDL针头(P<0.05);5)采血失败原因集中在血量不足和血液凝固,Capitainer采血设备搭配各针头使用失败率最高,HemaXis和HemaSpot采血设备搭配各针头失败率较低。结论:1)DBS技术采集血样时,建议使用用时更短、合格率更高的指尖血采集形式;2)采血针推荐使用BDM和BDH型号。3)结合采血器材、运动员偏好、疼痛感、采血合格率、采血时间等角度综合考虑,不推荐Capitainer采血器材,HemaXis采血器材推荐搭配BDM或BDH采血针使用,HemaSpot采血器材可搭配SL、SM、SH、BDM、BDH采血针使用。

液相色谱-串联质谱法检测干血点中的同型半胱氨酸

建立了一种高通量液相色谱-串联质谱技术检测干血点(DBS)中同型半胱氨酸(homocycteine,Hcy)的方法。以DBS为样本,homocystine-D8为同位素内标,二硫苏糖醇(DTT)为蛋白结合态Hcy的还原剂,使用含0.1%(v/v)甲酸、0.05%(v/v)三氟乙酸的乙腈溶液萃取。整个前处理过程使用自动移液平台及96孔板实现高通量自动化操作。处理后的样本经过Phenomenex CN柱分离,使用多反应监测模式进行LC-MS/MS分析。结果表明:Hcy的检出限为0.12μmol/L(S/N=3),定量限为0.46μmol/L(S/N=10)。Hcy在1.16~148.00μmol/L范围内线性关系良好,R^2=0.994。Hcy的平均回收率为(103.0±4.97)%^(112.0±2.13)%,日内相对标准偏差(RSD)为1.9%~4.6%,日间RSD为1.5%~7.1%。DBS样本在不同温度(-4、-20、22和37℃)下储存不同时间(0、1、2、3、4、5、6、14天)后的稳定性试验显示样本总体RSD<15%,经前处理后的样本在48 h内的稳定性试验显示样本总体RSD<5%。该方法与传统生化分析方法的相关性好(R^2=0.9818,n=47)。

液相色谱-串联质谱法快速检测干血点样品中尿酸

建立了一种高通量液相色谱-串联质谱(LC-MS/MS)检测干血点(DBS)样品中尿酸(UA)的方法。采用自动液体操作平台对样品进行高通量自动化前处理,首先用含有UA-1,3-15 N2稳定同位素内标的Tris水溶液进行萃取,然后用含有0.1%甲酸、0.05%三氟乙酸的乙腈溶液沉淀蛋白质。处理后的样品经CN色谱柱分离,多反应监测(MRM)模式进行LC-MS/MS分析。结果表明,在DBS样品中,UA在7.8~1 000μmol/L浓度范围内的线性关系良好(R2=0.999);检出限为3.1μmol/L(S/N=3);定量限为12.5μmol/L(S/N=10);平均回收率为95%~101%;日内相对标准偏差(RSD)为4.2%~12%;日间RSD为5.3%~14%。以样品中UA检测结果的总体RSD不超过15%来考察样品稳定性,分别将样品在-20℃保持30天、在37℃保持7天、反复冻融5次,样品中UA检测结果总体RSD小于10%,表明样品稳定性良好。将该方法与传统生化分析方法相比较,并分析了204份血样,相关性较好(R2=0.946)。此方法可为有限采血条件下UA的检测及UA相关疾病的大规模筛查提供新途径。

Evaluation of the AGCU Expressmarker 16 and 22 PCR Amplification Kits Using Biological Samples Applied to FTA Micro Cards in Reduced Volume Direct PCR Amplification Reactions

This study evaluated the performance of the Wuxi AGCU ScienTech Incorporation(HuiShan,Wuxi,China)AGCU Expressmarker 16(EX 16)and 22(EX22)short tandem repeat(STR)amplification kits in reduced reaction volumes using direct polymerase chain reaction(PCR)amplification workflows.The commercially available PowerPlex21(PP21)System(Promega,Wisconsin,USA),which follows similar direct workflows,was used as a reference.Anticoagulate blood applied to chemically impregnated FTATM Micro Cards(GE Healthcare UK Limited,Amersham Place,Little Chalfont,Buckinghamshire,HP79NA,UK)was used to represent a complex biological sample.Allelic concordance,first‑pass success rate,average peak heights,heterozygous peak height ratios(HPHRs),and intracolor and intercolor peak height balance were determined.In reduced volume PCR reactions,the performances of both the EX16 and EX22 STR amplification kits were comparable to that of the PP21 System.The level of performance was maintained at PCR reaction volumes,which are 40%of that recommended.The EX22 and PP21 System kits possess comparable overlapping genome coverage.This study evaluated the performance of the AGCU EX16 and EX22 STR amplification kits in reduced PCR reaction volumes using direct workflows in combination with whole blood applied to FTATM Micro Cards.Allelic concordance,first‑pass success rate,average peak heights,HPHRs,and intracolor and intercolor peak height balance were determined.A concordance analysis was completed that compared the performance of the EX16 and EX22 kits using human blood applied to FTA Micro Cards in combination with full,half,and reduced PCR reaction volumes.The PP21 System(Promega)was used as a reference kit.Where appropriate,the distributions of data were assessed using the Shapiro‑Wilk test.For normally‑distributed data,statistics were calculated using analysis of variance(ANOVA)and for nonparametric data the Wilcoxon/Kruskal‑Wallis test was used.Statistical significance was set at P<0.05.Confidence intervals for mean values were set at 95%.On using reduced volume PCR reactions in combination with dried blood spots applied to FTA sample collection cards,both the EX16 and EX22 kits were shown to generate STR profiles of sufficient quality to allow entry into National DNA databases.The performance of both EX16 and EX22 was comparable to that of the PP21 System.This study demonstrates the successful use of the Wuxi AGCU ScienTech Incorporation EX16 and EX22 kits in reduced PCR reaction volumes with complex biological samples applied to chemically impregnated FTA sample collection cards.

采用干血斑进行丙肝病毒RNA检测的研究

目的用干血斑（dried blood spot，DBS）取代静脉血作为检测HCV RNA的样品来源的可行性。方法评价该方法的检测限、批间和批内变异。用HCV RNA阴性的血液将第二代HCV RNA国际标准（WHO，HPAUK）稀释后浓度分别为1000，500，250和150IU／ml的样品检测。批间变异是通过反复检查单个1000IU／mlDBS对照超过20次PCR反应。批内变异则是通过一次PCR反应，在20个反应孔检测1000IU／ml对照DBS。从单个已知血液样品HCV阳性的标本制备成DBS后在不同温度和时间进行评估检测HCVRNA的影响，如室温下，4℃，-20℃和-80℃。另外在不同时间点检测，如1，2，4，6，9和12个月。结果DBS样品检测HCVRNA的检测限为250IU／ml。1000IU／ml样本的批间和批内变异系数（CV）分别为2．5％和2．2％。灵敏度和特异度分别为100％和95．8％。在不同温度保存超一年的DBS样品中HCVRNA是稳定的。结论笔者建立了一种灵敏和稳定的采用DBS检测HCVRNA的方法。用干血斑作为一种样品有利于对高危人群HCVRNA的检测。

Early Infant Diagnosis (EID) of HIV: An Experience at a Tertiary Care Hospital in India

Introduction: Early infant diagnosis (EID) confers substantial benefit to HIV infected and HIV uninfected infants and to programmes providing prevention of mother to child transmission (MTCT), but has been challenging to implement in resource limited settings. Objectives: To find out the rate of perinatal transmission in infants born to HIV positive mothers, to study the effect of various predisposing factors on HIV transmission and to evaluate the utility of dried blood spot (DBS) specimen for EID of HIV. Methods: Infants born to HIV positive mothers were tested according to National AIDS Control Organization (NACO) guidelines. Infants of 6 weeks to 6 months of age (n = 84) were diagnosed by DBS PCR;DBS positive results were confirmed by whole blood PCR. Infants 6 - 18 months (n = 47) were subjected to antibody test and if positive were confirmed by DNA PCR. Detailed history including type of delivery, single dose nevirapine (SDN) and breast feeding was taken. Results: The HIV transmission rate was 10.69%. In children ≤ 6 months, who did not receive SDN the positivity was 44.44% (4/9) whereas in those who received SDN it was 6.66% (5/75), (P = 0.0063). In children > 6 months the positivity rate was significantly higher in breast fed 42.85% (3/7) as compared to non breast fed 5% (2/40) children (P = 0.0187). There was 100% concordance between DBS and whole blood PCR. Conclusions: In resource limited settings, though HAART should be considered to further reduce MTCT during pregnancy and to prevent the emergence of resistance, SDN should be kept as an option for mothers coming directly in labour. Also, extended ART should be provided to mothers who want to breast feed their children. Early infant diagnosis using DBS specimens will further reduce the morbidity and mortality in these children.

Early infant diagnosis of HIV in India—Early results and sero-positivity determinants

Introduction: PMTCT under NACP-III cover ex-posed children born to sero-positive mothers. Baby’s sero-status could be confirmed only at 18 months. Under EID, by DBS and/or WB collection, DNA-PCR can be performed earlier, with subsequent ART-linkage and 18-months-con- firmation. In Ahmedabad, with 55,000 annual pre- gnancy-HIV-testing, sero-prevalence is 0.27%. Methodology: Entry-points in EID are at 6 weeks, 6 months or 12 months. Cohort of 213 exposed children since EID roll-out (June 2010-December 2011) at all tertiary care hospitals under Ahme-dabad Municipal Corporation was assessed for sero-positivity-prevalence, DBS validity and assessment of baby’s sero-status-determinants. De-identified, secondary data were captured under routine public-health-program. Necessary permissions taken. Results: 144 HIV sero-positive deliveries took place. 213 exposed children were enrolled in EID. Cumulatively, 18 (8.45%) were tested positive at all entry-points. Out of sero-positives confirmed at 18 months, 60% children’s mothers were detected either in second or third trimester. In 40%, mothers remained undiagnosed intra-partum. Mothers were not on ART intra-partum in 80% (RR 1.8). Peri-partum ARV prophylaxis-single-dose-Nevirapine (sdNVP) was not given in 60%. [RR 18, CI 3.69 to 87.70 at 95% (p < 0.0003)]. In 60%, mode of delivery was vaginal, deliveries were handled in emergency. History of exclusive breastfeeding was in 60%. Discussion: Rise in yield of sero-positivity with age, highest proportion of sero-positivity and highest number of entrants at 6 weeks call for efforts targeted towards increasing earliest EID uptake clubbed with immunization visits. Feasibility, validity and early-ART-linkage to reduce mortality are features of DBS. Results justify its use in national program. Earliest pregnancy-HIV detection, HIV-testing for emergency deliveries, intra-partum sdNVP to both mother and baby, ART-linkage of eligible mothers and following infant feeding guidelines remain cornerstone of PMTCT success.

自助采集尿液及干血斑样本检测HIV的结果评价

目的通过对一种艾滋病病毒(HIV)自助采样包采集的样本进行检测,探讨其临床应用的可行性。方法针对已知感染状态的男男性行为者(MSM)、静脉吸毒者(IDUs)和健康体检人群,共807人;未知感染状态的暗娼283人;由专业人员分发并回收HIV的自助采样包。利用酶联免疫吸附试验对收集的尿液及干血斑样本进行HIV抗体检测,并对暗娼进行干血斑核酸检测,分析不同方法检测结果的一致性。结果尿液样本抗体检测结果与已知感染状态比较,健康体检人群、MSM、IDUs以及总人群的检测一致率分别为96.55%、98.99%、95.20%、97.03%;干血斑样本抗体检测结果与已知感染状态比较,健康体检人群、MSM、IDUs以及总人群的检测一致率分别为98.47%、100%、98.80%、99.13%。对未知感染状态的暗娼,将抗体检测结果与干血斑核酸检测结果进行比较,干血斑样本与尿液样本的一致率分别为96.82%、97.88%;将干血斑与尿液样本的抗体检测结果比较,检测一致率为96.82%。结论对采用自助采样包采集的样本进行检测,发现具有较高的检测效能,说明自助采样包采集的样本,不仅适用于高危人群,在自然人群中也有实用价值。

HIV-1母婴传播早期诊断检测方法应用

目的应用核酸扩增技术对母婴传播婴幼儿人类免疫缺陷病毒（HIV）感染进行早期诊断,为临床应用提供依据。方法对2009-2010年HIV阳性孕产妇活产的34例3-14个月龄婴幼儿收集足底干血斑样本进行HIV-1 DNA检测。随访18个月后进行血清HIV抗体检测比较。结果 34例婴幼儿中,检出HIV-1 DNA 9例,检出率26.5%。9例HIV-1 DNA阳性患儿均未使用过抗病毒治疗。18个月后随访,34例婴幼儿除1例失访,2例检出HIV-1 DNA婴儿已死亡（根据临床有机会性感染体征诊断为AIDS）,7例检出HIV-1 DNA婴儿血清抗体阳性外,其他24例未检出HIV-1 DNA的婴儿其血清抗体为阴性。两种方法检测结果比较符合率为100.0%。结论核酸检测是有效的早期诊断方法,早期诊断有利于后续开展母婴阻断工作。

LC-MS/MS法测定干血斑样品中阿戈美拉汀

目的：建立快速、灵敏的液相色谱-串联质谱（LC-MS/MS）法,测定大鼠干血斑（DBS）样品中的阿戈美拉汀,并应用于阿戈美拉汀大鼠药动学研究。方法：给药后的大鼠新鲜全血40μL点于Whatman 903滤纸上,室温下干燥2 h,放入含干燥剂的密封袋保存。取直径为6 mm的干血斑3片,加入甲醇500μL,超声15 min后,进行液-液萃取,以甲醇-甲酸铵-甲酸为流动相,流速0.20 m L·min-1,采用Grom-Sil 120 ODS-5 ST柱分离,通过电喷雾三重四极杆串联质谱,以多反应监测（MRM）方式进行检测,用于定量的离子反应为m/z 244.1→m/z 185.1（阿戈美拉汀）和m/z 249.2→m/z 187.1（d5-阿戈美拉汀）。结果：测定阿戈美拉汀DBS的线性范围为0.025 0-25.0 ng·m L-1;定量下限为0.025 0 ng·m L-1;精密度和准确度均符合要求。结论：DBS采样法全血用量少,样品储存、运输便利,可用于阿戈美拉汀大鼠药动学研究。

新疆南疆地区2013年-2015年新生儿疾病筛查血片质量情况分析

目的分析新疆南疆5市37区县2013年-2015年新生儿疾病筛查血片质量整体情况,探讨南疆地区近三年新生儿疾病筛查不合格血片的原因。方法根据《新生儿疾病筛查血片采集技术规范》对新疆南疆地区新生儿疾病筛查的血片质量进行分析,统计不合格血片并分析原因。结果新疆南疆地区近三年共筛查新生儿418 469例,平均筛查率为46.47%;首次采集血片不合格数19 709例,平均不合格率为4.56%;首次补采血片4856例,平均首次补采率为24.64%;集中统计为六类不合格因素,最主要原因是血片污染。结论采集血片是新生儿疾病筛查重要部分,血片质量是否合格将直接影响新生儿疾病筛查检测结果的准确性。提高血片质量对新生儿疾病筛查有重要的意义。

干血点采样技术和干血浆采样技术在麦考酚酸药动学分析中的应用

目的探讨干血点采样技术(dried blood spots,DBS)和干血浆采样技术(dried plasma spots,DPS)在麦考酚酸药动学分析中的应用。方法色谱柱为Ecosil C18(150mm×4.6mm,5μm),流动相为0.01moL/L甲酸水(0.1%甲酸)-乙腈-甲醇(25:45:30),体积流量为0.4mL/min,柱温为30℃,进样量10μL;质谱检测采用ESI离子源,正离子MRM方式检测;考察可能的影响因素并进行临床研究。结果 DBS法和DPS法方法学和临床实验数据均良好。结论 DBS方法和DPS法较一般处理方法简便,用血量少,储存运输方便,在一些药物的药动学分析中能代替一般的处理方法。

干血点采样技术及其在药动学研究中的应用

干血点采样(dried blood spot,DBS),即将全血样品收集在卡纸上,在血样采集方法上比传统方法有一定的优势。它需求较少的血量,可减少动物使用,方便血样采集、存储运输,简化样品前处理。到目前为止,DBS与LC-MS/MS技术联用在小分子定量分析中已成为一个重要的方法,将在药动学研究中得到越来越多的应用。概述DBS样品收集,储存、前处理以及分析方法。

干血点技术在滥用药物分析中的应用研究进展

干血点(DBS)作为一种新型的采血技术被广泛应用于临床以及人体中重要物质的测定等领域,与此同时在毒品分析领域中的应用也不断增加。DBS技术具有需血量少、操作简单、稳定性强和易于保存运输等优点,完美契合了毒品分析中对检材处理的需求。与全血分析相比,DBS与液质联用等检测方法结合对部分常见毒品的分析结果展现出了独特的优势。本文对DBS采样技术在滥用药物分析中的应用、难点、解决措施和发展前景进行了综述,旨在为法庭科学中深入研究和开发DBS技术的应用提供参考。