Contamination of dried blood spots – an underestimated risk in newborn screening

2018 ◽  
Vol 56 (2) ◽  
pp. 278-284 ◽  
Author(s):  
Theresa Winter ◽  
Anja Lange ◽  
Anke Hannemann ◽  
Matthias Nauck ◽  
Cornelia Müller
Keyword(s):  
Newborn Screening ◽  
Infant Formula ◽  
False Positive ◽  
Dried Blood Spots ◽  
Substantial Effect ◽  
Blood Collection ◽  
Inborn Errors ◽  
Blood Spots ◽  
Filter Papers ◽  
The Impact

Abstract Background: Newborn screening (NBS) is an established screening procedure in many countries worldwide, aiming at the early detection of inborn errors of metabolism. For decades, dried blood spots have been the standard specimen for NBS. The procedure of blood collection is well described and standardized and includes many critical pre-analytical steps. We examined the impact of contamination of some anticipated common substances on NBS results obtained from dry spot samples. This possible pre-analytical source of uncertainty has been poorly examined in the past. Methods: Capillary blood was obtained from 15 adult volunteers and applied to 10 screening filter papers per volunteer. Nine filter papers were contaminated without visible trace. The contaminants were baby diaper rash cream, baby wet wipes, disinfectant, liquid infant formula, liquid infant formula hypoallergenic (HA), ultrasonic gel, breast milk, feces, and urine. The differences between control and contaminated samples were evaluated for 45 NBS quantities. We estimated if the contaminations might lead to false-positive NBS results. Results: Eight of nine investigated contaminants significantly altered NBS analyte concentrations and potentially caused false-positive screening outcomes. A contamination with feces was most influential, affecting 24 of 45 tested analytes followed by liquid infant formula (HA) and urine, affecting 19 and 13 of 45 analytes, respectively. Conclusions: A contamination of filter paper samples can have a substantial effect on the NBS results. Our results underline the importance of good pre-analytical training to make the staff aware of the threat and ensure reliable screening results.

scholarly journals EDTA in Dried Blood Spots Leads to False Results in Neonatal Endocrinologic Screening

2008 ◽  
Vol 54 (3) ◽  
pp. 602-605 ◽  
Author(s):  
Ute Holtkamp ◽  
Jeanette Klein ◽  
Johannes Sander ◽  
Michael Peter ◽  
Nils Janzen ◽  
...  
Keyword(s):  
Newborn Screening ◽  
False Positive ◽  
Neonatal Screening ◽  
False Negative ◽  
Dried Blood Spots ◽  
Screening Tests ◽  
Blood Collection ◽  
Blood Spots ◽  
Sampling Procedures ◽  
Positive Results

Abstract Background: Blood samples for neonatal screening for inborn errors of metabolism are collected and shipped on standardized filter paper cards. Occasionally these samples are contaminated with EDTA, which is often used for anticoagulation. EDTA may interfere with newborn screening tests based on lanthanide fluorescence and thus lead to false-negative or false-positive results. Methods: We used tandem mass spectrometry (MS/MS) to detect EDTA in dried blood spots by use of an extra experiment that was integrated into the standard MS/MS neonatal screening and did not require an additional sample spot, nor extra time or work. We analyzed the influence of different blood sampling procedures on lanthanide fluorescence tests for thyroid-stimulating hormone (TSH) and 17-hydroxyprogesterone (17-OHP). Results: EDTA was increased in 138 of 190 000 newborn screening samples, 27 of which caused false- positive results in the immunoassay for 17-OHP. No false-negative TSH results were found. False-positive results in the 17-OHP test occurred when EDTA concentrations were >2.0 g/L; the TSH test, however, produced false negatives only when EDTA concentrations were >3.0 g/L. Using EDTA-containing devices the procedure of blood collection significantly influenced the concentration of the anticoagulant. Conclusion: Addition of EDTA quantification into standard MS/MS tests is a simple and useful method to avoid false-positive or false-negative neonatal screening results in lanthanide fluorescence–based tests.

scholarly journals The Impact of Seasonal Changes on Thyroxine and Thyroid-Stimulating Hormone in Newborns

2021 ◽  
Vol 7 (1) ◽  
pp. 8
Author(s):  
Rebecca McMahon ◽  
Lenore DeMartino ◽  
Mycroft Sowizral ◽  
Diana Powers ◽  
Melissa Tracy ◽  
...  
Keyword(s):  
Newborn Screening ◽  
False Positive ◽  
Seasonal Changes ◽  
Dried Blood Spots ◽  
Thyroid Stimulating Hormone ◽  
Seasonal Temperature ◽  
Blood Spots ◽  
Significant Difference ◽  
The Impact ◽  
Stimulating Hormone

Newborn screening for congenital hypothyroidism (CH) is performed by measuring the concentration of thyroxine (T4) and/or thyroid-stimulating hormone (TSH) in dried blood spots. Unfortunately, the levels of T4 and TSH vary due to multiple factors, and therefore the false-positive rate for the test is a challenge. We analyzed screening data from 2008 to 2017 to determine the effect of seasonal changes and manufacturer kit lot changes on T4 and TSH values and on numbers of infants referred. Over a 10-year period, we screened 2.4 million infants using commercially available fluoroimmunoassays to measure T4 and TSH concentrations in dried blood spots. During colder months, daily mean T4 and TSH values were higher and referral rates and false-positive rates were higher. However, there was no significant difference between the number of confirmed CH cases. Furthermore, in rare instances, we observed differences in T4 daily mean values during the 10-year period when manufacturer kit lot changes were made. Seasonal temperature variations influence measured T4 and TSH values and consequently lower the positive predictive value for CH testing in colder months. Newborn screening (NBS) programs should be aware that manufacturer kit lot changes may also influence T4 values.

scholarly journals NeoSeq: a new method of genomic sequencing for newborn screening

2021 ◽  
Vol 16 (1) ◽  
Author(s):  
Huaiyan Wang ◽  
Yuqi Yang ◽  
Lingna Zhou ◽  
Yu Wang ◽  
Wei Long ◽  
...  
Keyword(s):  
Newborn Screening ◽  
False Positive ◽  
Disease Risk ◽  
Amplicon Sequencing ◽  
Dried Blood Spots ◽  
Inborn Errors ◽  
Screening Cost ◽  
Diagnostic Time ◽  
Significant Difference ◽  
Positive Results

Abstract Objective To explore the clinical application of NeoSeq in newborn screening. Methods Based on the results obtained from traditional newborn screening (NBS) with tandem mass spectrometry (TMS), three cohorts were recruited into the present study: 36 true positive cases (TPC), 60 false-positive cases (FPC), and 100 negative cases. The dried blood spots of the infants were analyzed with NeoSeq, which is based on multiplex PCR amplicon sequencing. Results Overall, the sensitivity of NeoSeq was 55.6% (20/36) in the detection of TPC. NeoSeq detected disease-related genes in 20 of 36 TPC infants, while it could not identify these genes in eight children. Five cases (3.1%) with disease risk were additionally found in the FPC and NC cohorts. There was a significant difference in the diagnostic time between the two methods—10 days for NeoSeq vs. 43 days for traditional NBS. Conclusions NeoSeq is an economic genomic screening test for newborn screening. It can detect most inborn errors of metabolism, reduce the rate of false positive results, shorten the porting cycles, and reduce the screening cost. However, it is still necessary to further optimize the panel design and add more clinically relevant genomic variants to increase its sensitivity.

scholarly journals A False-Positive Case of Methylmalonic Aciduria by Tandem Mass Spectrometry Newborn Screening Dependent on Maternal Malnutrition in Pregnancy

2020 ◽  
Vol 17 (10) ◽  
pp. 3601 ◽  
Author(s):  
Claudia Rossi ◽  
Ilaria Cicalini ◽  
Cristiano Rizzo ◽  
Mirco Zucchelli ◽  
Ada Consalvo ◽  
...  
Keyword(s):  
Newborn Screening ◽  
False Positive ◽  
Vitamin B12 Deficiency ◽  
Propionic Acidemia ◽  
Dried Blood Spots ◽  
Specific Marker ◽  
Inborn Errors ◽  
Maternal Malnutrition ◽  
Ms Analysis ◽  
Second Tier

Methylmalonic Acidurias (MMAs) are a group of inborn errors of metabolism (IEMs), specifically of propionate catabolism characterized by gastrointestinal and neurometabolic manifestations resulting from a deficiency in the function of methylmalonyl-CoA mutase, methylmalonyl-CoA epimerase, and cobalamin metabolism. In Expanded Newborn Screening (NBS), increased levels of propionylcarnitine (C3) and/or of its ratios by MS/MS analysis of dried blood spots (DBS) samples are suggestive for either Propionic Acidemia or MMAs. C3 elevation is not considered a specific marker for these disorders, resulting in high false-positive rates. The use of analyte ratios improves specificity, but it still cannot resolve the diagnostic issue. Second-tier testing are strongly recommended as confirmation of primary NBS results and for a differential diagnosis. LC-MS/MS analysis allows the quantification of more specific markers of the disorder. Here, we report the case of a newborn with a suspected MMA at Expanded NBS and at second-tier test. Given the urgent situation, in-depth diagnostic investigations were performed. Further investigations surprisingly revealed a Vitamin B12 deficiency due to a maternal malnutrition during pregnancy. This case emphasized that metabolic alterations at NBS may not only be influenced by genome and related to IEMs, but also to external factors and to maternal conditions.

The Acylcarnitine Profile in Dried Blood Spots is Affected by Hematocrit: A Study of Newborn Screening Samples in Very-Low-Birth-Weight Infants

2020 ◽  
Author(s):  
Mitsuhiro Haga ◽  
Mitsuhisa Isobe ◽  
Ken Kawabata ◽  
Masaki Shimizu ◽  
Hiroshi Mochizuki
Keyword(s):  
Birth Weight ◽  
Low Birth Weight ◽  
Newborn Screening ◽  
Very Low Birth Weight ◽  
Dried Blood Spots ◽  
Screening Tests ◽  
Low Birth Weight Infants ◽  
Inborn Errors ◽  
Blood Spots ◽  
Acylcarnitine Profile

Objective The acylcarnitine profile is analyzed in dried blood spots (DBS) to screen for inborn errors of metabolism. Hematocrit (Ht) is known to affect the result of quantitative analyses of DBS samples; however, the effects of Ht on the acylcarnitine profiles in DBS have not been studied in actual samples from newborns. Study Design The acylcarnitine profiles in DBS for newborn screening tests and Ht levels of very-low-birth-weight infants were obtained from medical records. We investigated the relationship between Ht and each acylcarnitine using Pearson's correlation coefficient (r). Results We examined 77 newborns in this study. There was a significantly positive correlation between Ht and C0, C2, C12, C16, C18, C18:1, and C18:1-OH, respectively (p < 0.0025). The correlation was the greatest on C2 (r = 0.59). Conclusion This study clarifies that Ht and C0, C2, C12, C16, C18, C18:1, and C18:1-OH are significantly correlated in DBS, which is consistent with previous studies. Hence, the effect of Ht should be considered when interpreting the results of acylcarnitine profiles in DBS. Key Points

scholarly journals Diagnostic potential of stored dried blood spots for inborn errors of metabolism: a metabolic autopsy of medium-chain acyl-CoA dehydrogenase deficiency

2018 ◽  
Vol 71 (10) ◽  
pp. 885-889 ◽  
Author(s):  
Noriyuki Kaku ◽  
Kenji Ihara ◽  
Yuichiro Hirata ◽  
Kenji Yamada ◽  
Sooyoung Lee ◽  
...  
Keyword(s):  
Newborn Screening ◽  
Inborn Errors Of Metabolism ◽  
Dried Blood Spots ◽  
Screening Tests ◽  
Inborn Errors ◽  
Mcad Deficiency ◽  
Medium Chain ◽  
Blood Spots ◽  
Chain Acyl ◽  
Biochemical Analyses

AimIt is estimated that 1–5% of sudden infant death syndrome (SIDS) cases might be caused by undiagnosed inborn errors of metabolism (IEMs); however, the postmortem identification of IEMs remains difficult. This study aimed to evaluate the usefulness of dried blood spots (DBSs) stored after newborn screening tests as a metabolic autopsy to determine the causes of death in infants and children who died suddenly and unexpectedly.MethodsInfants or toddlers who had suddenly died without a definite diagnosis between July 2008 and December 2012 at Kyushu University Hospital in Japan were enrolled in this study. Their Guthrie cards, which had been stored for several years at 4–8°C, were used for an acylcarnitine analysis by tandem mass spectrometry to identify inborn errors of metabolism.ResultsFifteen infants and children who died at less than 2 years of age and for whom the cause of death was unknown were enrolled for the study. After correcting the C0 and C8 values assuming the hydrolysation of acylcarnitine in the stored DBSs, the corrected C8 value of one case just exceeded the cut-off level for medium-chain acyl-CoA dehydrogenase (MCAD) deficiency screening. Genetic and biochemical analyses confirmed this patient to have MCAD deficiency.ConclusionDBSs stored after newborn screening tests are a promising tool for metabolic autopsy. The appropriate compensation of acylcarnitine data and subsequent genetic and biochemical analyses are essential for the postmortem diagnosis of inborn errors of metabolism.

scholarly journals NeoSeq: A New Method of Genomic Sequencing for Newborn Screening

2021 ◽  
Author(s):  
huaiyan wang ◽  
yuqi yang ◽  
lignna zhou ◽  
yu wang ◽  
wei long ◽  
...  
Keyword(s):  
Newborn Screening ◽  
False Positive ◽  
Disease Risk ◽  
Amplicon Sequencing ◽  
Dried Blood Spots ◽  
Screening Methods ◽  
Genomic Sequencing ◽  
Inborn Errors ◽  
Screening Cost ◽  
Positive Results

Abstract Objective To explore the clinical application of NeoSeq in newborn screening. Methods Based on the results obtained from traditional newborn screening (NBS), three cohorts were recruited into the present study: 36 true positive cases (TPC), 60 false-positive cases (FPC), and 100 negative cases. The dried blood spots of the infants were analyzed with NeoSeq, which is based on multiplex PCR amplicon sequencing. Results Overall, the sensitivity of both NeoSeq and traditional NBS projects was 55.6% (20/36) in the detection of TPC. NeoSeq detected disease-related genes in 20 of 36 TPC infants, while it could not identify these genes in eight children. Five cases (3.1%) with disease risk were found in the FPC and NC cohorts. There was a significant time difference in the diagnostic result cycles between the two methods − 10 days for NeoSeq vs. 43 days for traditional NBS. Conclusions NeoSeq is an effective method of genomic sequencing for newborn screening. It can detect most inborn errors of metabolism, reduce the rate of false positive results, shorten the reporting cycles, and reduce the screening cost.

scholarly journals QUALITY OF DRIED BLOOD SPOTS IS AN INTEGRAL COMPONENT OF PROMPT DETECTION OF INBORN ERRORS OF METABOLISM

2020 ◽  
Vol 10 (4(38)) ◽  
pp. 77-86
Author(s):  
Tetiana Znamenska ◽  
O. Vorobiova ◽  
I. Kuzneczov ◽  
I. Lastivka ◽  
A. Kremezna ◽  
...  
Keyword(s):  
Newborn Screening ◽  
Inborn Errors Of Metabolism ◽  
Dried Blood Spots ◽  
Poor Quality ◽  
Blood Sampling ◽  
Inborn Errors ◽  
Blood Spots ◽  
Expanded Newborn Screening ◽  
Practical Recommendations

Introduction. Inborn Errors of Metabolism (IEM) are constituted a group of genetic diseases that are associated with defects in the synthesis or catabolism of complex molecules, impaired intermediary metabolism and energy production/utilization processes. The clinical manifestation of IEM is nonspecific, that looks similar to septicemia, and most often occurs in the neonatal period with life-threatening acute metabolic crises. Expanded Newborn Screening (ENS) – a biochemical study of the blood of all newborns without exception with the purpose to identify molecular markers of these diseases proved to be the most effective instrument of early IEM diagnostics. The quality of the biological samples (dried blood spots, DBS) in great extent determines the timing, accuracy, and reliability of the results of biochemical measurements. Obtaining of equivocal results in the case of analysis of poor quality DBS requires repeated laboratory tests, that delays the diagnostic process and postpones the start of specific treatment, which usually results in irreversible damage of the brain and internal organs of the child. The aim of this work is to (i) review the first results of the implementation of Expanded Newborn Screening in Ukraine (pilot part of the Baby Screen Project), and to analyze literature data regarding the negative impact of poor quality DBS on laboratory determination of IEM marker substances contents in the specimens, (ii) to characterize the typical errors in blood sampling and drying of  blood spots, and (iii) to provide practical recommendations for the proper performance of these procedures. Materials and methods. Own data of retrospective analysis of the questionable ENS results was superimposed with dried blood specimens, that were investigated in the Pharmbiotest ENS Lab to outline most common inaccuracies. Based on the comparison of these data with the relevant publications it was formulated the practical recommendations for improving quality of DBS preparation to ensure the accuracy and reliability of laboratory measurements and speed up IEM diagnostics. Results. The quality of biomaterial selection is an important part of obtaining reliable results during expanded newborn screening. Capillary blood is collected in the maternity hospital from 48-72 hours (full-term) and for 7-11 days (in preterm babies) after the birth from the heel of babies. In this case, a few drops of blood are applied to a special test card made of filter paper, which is dried and sent to the laboratory. Blood tests are performed using a highly sensitive and accurate method of chemical analysis - tandem mass spectrometry in the laboratory "Pharmbiotest", located in Ukraine. Taking into analysis the low-quality samples lead to questionable results, which requires repeated DBS sampling and re-examining. This proved to be the most common cause of delaying IEM detection, diagnosis establishment, and initiation of treatment, which can be fatal for a child with severe IEM forms. Conclusions. Informing healthcare professionals and parents about the current results of laboratory monitoring of dried blood spots quality, typical errors in blood sampling and following on-site procedures and negative consequences of its improper performance, as well as providing clear practical recommendations of how these procedures should be done is a proven way of improving and speeding up IEM diagnostics.

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