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.

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.

The Use of Dried Blood Spot Samples in Screening Drugs of Abuse

The present article will provide an overview of use of dried blood sampling method for analysis of drug of abuse. Relatively short half life and instability in blood, calls for alternative sampling method for determination of drugs of abuse. Dried blood spot (DBS) method has many advantages over the conventional sampling methods. The available method for DBS sample collection, storage and transport are described here. The techniques involved in and the factors that may influence the accuracy and reproducibility of the DBS methods for determination of drugs of abuse are presented. The DBS sampling has the potential to be a useful technique to detect drugs of abuse. The use of DBS for any drug should be judged against the potential error involved with the method.

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.

Reliability of enzyme assays in dried blood spots for diagnosis of 4 lysosomal storage disorders

Introduction: Lysosomal storage disorders (LSD) are inherited diseases caused, in the majority of them, by the deficiency of lysosomal enzymatic activities. Ob-jectives: We aimed to analyze the usefulness of DBS samples for diagnosis of 4 LSDs, with the availability of a large quantity of patient samples. Design and methods: Blood samples from previously diagnosed patients with Fabry, Gaucher, Hunter, and Maro-teaux-Lamy syndromes and normal control individ-uals, were collected and dispen-sed in filter paper, and used for enzymatic activity determination. Results: Diagnosis of hemi/homo-zygous patients with Fabry, Hunter and Maroteaux-Lamy diseases using DBS samples showed ideal parameters of 100% sensitivity and specificity. DBS assay for Gaucher disease would need a posterior confirmatory step. Conclusions: Leukocyte measu-rement is the only reliable way to diagnose Gaucher disease. For Hunter, Fabry and Maroteaux-Lamy disorders discrimination between patients and controls seems adequate by DBS.

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.

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.

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.

干血斑中34种苯二氮卓类物质的实时直接分析-串联质谱快速筛查研究

该文通过优化仪器条件和前处理方法建立了实时直接分析-串联质谱快速筛查干血斑(DBS)中34种苯二氮卓类物质(BZDs)的方法。首先将50μL血液沉积在采血卡上形成DBS,DBS经溶剂充分提取后取上层溶液,使用DART 12Dip-It^(TM)模块进样5μL,在多反应监测模式下检测。结果表明:选用400℃作为载气加热器温度,以乙酸乙酯-水(体积比3∶1)混合溶剂提取DBS样品,所得响应强度较高且建立的筛查方法选择性良好,无延迟效应影响,除阿地唑仑、奥沙西泮和α-羟基三唑仑外,其他物质的检出限均在5~50 ng/mL。所建方法可在90.3%的阳性检材中成功筛选出BZDs物质,方法快速、简便、有效,适用于服药自杀等类案件中DBS检材的快速筛查检验。

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。

液相色谱-串联质谱法测定干血点中晕车药及代谢物

该文建立了干血点(DBS)中东莨菪碱、阿托品、茶碱、地芬尼多、苯海拉明及二苯甲酮的液相色谱-串联质谱检测方法。取20μL血液制备干血点,以甲醇-乙腈(体积比8∶2)为溶剂进行超声提取,采用Thermo Gold ODS(150 mm×2.1 mm,5μm)色谱柱进行分离,以甲醇-5 mmol/L乙酸铵溶液(含0.2%冰乙酸,pH 4.0)为流动相梯度洗脱,在正离子电离方式下采用多反应监测模式进行测定,以外标法定量。结果表明:干血点中6种目标物在一定的质量浓度范围内线性关系良好,相关系数(r^(2))均不小于0.9930,检出限为3~30 ng/mL,定量下限为10~100 ng/mL;在低、中、高3个加标水平下,6种目标物的平均回收率为88.8%~108%,日间和日内相对标准偏差均小于15%。该方法操作简便、快捷,准确度和灵敏度高,已成功应用于1例地芬尼多中毒者的检测,为晕车药中毒案例的测定提供了新方法。

干血斑保存温度对新生儿遗传代谢病筛查结果的影响

目的探讨血斑卡在不同储存温度条件下对筛查标志物检测结果的影响,为提高新生儿遗传代谢病筛查前样本质量提供参考依据。方法纳入宜宾市妇幼保健院新生儿100例,每例采集干血斑3份,分别放置于2~8℃冷藏、20~24℃常温及37℃孵箱三种不同温度,储存48小时后进行新生儿遗传代谢病筛查,对不同储存条件下的结果进行分析。结果TSH、Phe、17α-OHP及G-6-PD四项指标的在不同保存条件下的检测结果均存在差异(P<0.05)。以低温存储的样本作为参考,发现37℃储存样本检测结果的精密度、及与参考值的相关性相对于常温存储均有所下降。从筛查结果的角度,37℃样本检测中产生了大量的假阳性结果,尤其是在G-6-PD缺乏症的检测中,假阳性率达到了90%。结论样本保存不当可能影响检测精密度、增加检测假阳性率。干血斑样本应尽量在低温运输及存储,尤其应当避免暴露于37℃及以上的环境,避免因保存不当产生假阳性结果。

Optimization and application of a dried blood spot-based genetic screening method for thalassemia in Shenzhen newborns

Background To optimize and apply an approach suitable for large-scale neonatal thalassemia genetic screening in China,thalassemia genotypes were determined by polymerase chain reaction-reverse dot blot using DNA extracted from dried blood spots(DBS)obtained from newborn screening programs.Methods Firstly,the most suitable commercial DNA extraction kit for DBS was screened.Then,the appropriate amount of DBS required for the automated high-throughput DNA extraction system was evaluated.Finally,the thalassemia prevalence and genotype spectrum in Shenzhen were investigated in 2028 newborns using the optimized screening procedure.Results The Magentec extraction kit was best suited for the automated DBS DNA extraction system using eight 3-mm DBS discs.The neonatal thalassemia prevalence in Shenzhen was 9.12%;6.31%α-thalassemia,2.37%β-thalassemia,and 0.44%α-/β-thalassemia.Conclusions Genetic screening based on DBS can precisely identify the thalassemia genotypes.Bothα-andβ-thalassemia are widely distributed in Shenzhen newborns.Newborn genetic screening is important for establishing a comprehensive thalassemia prevention program and for public education.

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.

滤纸干血片6种标志物UPLC-MS/MS方法的建立和性能评价

目的建立可同时检测新生儿苯丙酮尿症(PKU)、先天性肾上腺皮质增生症(CAH)和先天性甲状腺功能减退症(CH)6种标志物的超高效液相色谱串联质谱(UPLC-MS/MS)方法。方法对滤纸干血片的处理条件进行优化,建立可同时检出17-羟孕酮(17-OHP)、雄烯二酮(ASD)、皮质醇(COR)、苯丙氨酸(PHE)、酪氨酸(TYR)和甲状腺素(T4)的UPLC-MS/MS方法并进行方法学评价(线性范围、准确度、灵敏度、精密度、参考物质分析)。收集正常新生儿滤纸干血片样本1000份、CH患儿滤纸干血片样本30份,PKU患儿滤纸干血片样本30份、CAH患儿滤纸干血片样本30份。对建立的UPLC-MS/MS方法进行验证,并初步建立6种标志物筛查的参考区间。结果UPLC-MS/MS检测PHE、TYR、17-OHP、ASD、COR、T4的线性范围分别为12.1~980.7μmol/L、3.7~298.0μmol/L、6.1~490.2 nmol/L、0.7~56.6 nmol/L、13.8~1117.3 nmol/L、32.2~2606.6 nmol/L,最低检测限分别为0.5μmol/L、0.7μmol/L、1.6 nmol/L、0.2 nmol/L、2.6 nmol/L、3.2 nmol/L。3个浓度水平质控品的日内精密度[变异系数(CV)]为2.2%~7.5%,日间精密度(CV)为2.3%~7.3%,加标回收率为92.5%~115.4%。国际新生儿筛查学会(ISNS)PHE和17-OHP标准物质5个浓度的检测值与标示值的相对偏差分别为1.78%~7.01%、0.70%~10.78%。相对于正常新生儿,CAH患儿17-OHP水平和(17-OHP+ASD)/COR比值升高(P<0.05),PKU患儿PHE水平和PHE/TYR比值均升高(P<0.05),CH患儿T4水平降低(P<0.05)。17-OHP和17-OHP联合(17-OHP+ASD)/COR比值筛查CAH的阳性率均为100%。PHE和PHE联合PHE/TYR比值筛查PKU的阳性率均为100%。T4筛查CH的阳性率为40%。结论建立了可同时检测PKU、CAH和CH 6种标志物的UPLC-MS/MS方法,该方法特异性强、快速、灵敏,缩短了样本分析时间,可作为新生儿疾病筛查的新方法。

VAMS与LC-MS/MS技术联合及其临床应用的研究进展

在临床采样中体积吸收微量采样(VAMS)法有采血量少、创伤小及便于运输等独特优势,液相色谱-串联质谱(LC-MS/MS)法检测具有特异性强、灵敏度高、高通量及样品用量少等优点,两种技术结合可推动家中采样,实现远程诊断,符合精准医学和个体化治疗的发展要求。该文主要介绍了VAMS技术及VAMS法与干血斑(DBS)法的比较,VAMS和LC-MS/MS技术的联合应用情况和VAMS样品的处理。

可溯源干血片样本类固醇激素标准曲线的建立

目的建立可溯源的干血片样本19种类固醇激素液相色谱-串联质谱(LC-MS/MS)标准曲线。方法收集乙二胺四乙酸抗凝新鲜全血,用0.9%NaCl溶液清洗血细胞,将清洗后的血细胞和无类固醇激素的血浆按0.55∶0.45比例混合,制备模拟全血。根据拟制备标准曲线浓度添加不同量类固醇激素至模拟全血样本中,制备不同浓度的类固醇激素标准曲线干血片。采用液相色谱-串联质谱(LC-MS/MS)检测各标准曲线干血片类固醇激素含量,分析其线性范围,并对类固醇激素标准曲线干血片进行性能验证。结果类固醇激素标准曲线干血片中孕烯醇酮、孕酮、可的松、脱氢表雄酮的线性范围为0.25~100 ng/mL,11-脱氧皮质酮、21-脱氧皮质醇、18-羟皮质醇、11-酮睾酮的线性范围为0.05~20 ng/mL,皮质酮、11-羟睾酮的线性范围为0.025~10 ng/mL,17-羟孕酮、18-羟皮质酮、11-脱氧皮质醇、雄烯二酮、11-羟雄烯二酮、睾酮的线性范围为0.1~40 ng/mL,皮质醇的线性范围为1.5~600 ng/mL,17-羟孕烯醇酮的线性范围为0.2~80 ng/mL,硫酸脱氢表雄酮的线性范围为25~10000 ng/mL;相关系数(r^(2))≥0.997。结论建立的干血片样本类固醇激素标准曲线具有良好的线性,可实现相关项目测量结果的溯源。

滤纸干血滴抽提恶性疟原虫 DNA 用于 PCR 扩增

以ＳＴＥ－蛋白酶Ｋ－ＳＤＳ、甲醇－蛋白酶Ｋ－ＳＤＳ、Ｃｈｅｌｅｘ－１００加热法和Ｃｈｅｌｅｘ－１００蛋白酶Ｋ等四种方法对恶性疟患者滤纸干血滴样品进行ＤＮＡ抽提，供ＰＣＲ扩增特异性ＡＭＡ－１ＤＮＡ片段。结果显示，除ＳＴＥ－蛋白酶Ｋ－ＳＤＳ法抽提ＤＮＡ进行ＰＣＲ试验未能获得扩增产物外，其他３种方法抽提ＤＮＡ后进行ＰＣＲ试验均获得特异性约９００ｂｐＤＮＡ片段，可供进一步试验。