Screening Test For Deficiency Research Articles

Calculated globulin can be used as a screening test for antibody deficiency in children and adolescents

PurposeCalculated globulin (CG, total protein minus albumin levels) correlate well with IgG levels and has been proposed as a suitable screening method for individuals with primary antibody deficiencies (PADs). We aimed to show the correlation of CG with IgG levels in children and adolescents, utilizing a common method for albumin measurement, bromocresol green.MethodsIndividuals from two Allergy and Immunology clinics were invited to participate. Inclusion criteria were age < 18, stable conditions, and signed informed consent. We included 1084 individuals. Immunoglobulin G values were determined by immunoturbidimetry; the colorimetric bromocresol green method and the Architect Biuret method were utilized for the albumin and total protein (TP) measurements, respectively.ResultsA total of 1084 individuals were included in the analysis and divided into 4 age groups (0 to <1 year, 1 to <4 years, 4 to <10 years, and 10 to <18 years). For all patients, the mean age was 6.1 (± 5) years old, the mean IgG was 9.4 (± 4.7) g/L, and CG was 23.7 (± 5.9) g/L. The most frequent diagnosis were respiratory allergies, followed by inborn errors of immunity. IgG and CG varied according to age group. Cutoff values for hypogammaglobulinemia varied from 13.8 g/L in children < 1 year to 23.1 g/L in children and adolescents aged 10 to <18 years. CG sensitivity varied from 70.9% in children aged 1 to <4 years old to 95.8% in children 4 to <10. Specificity ranged from 87.5% in children 4 to <10 years old to 100% in children and adolescents aged 10 to <18 years.ConclusionCG is a suitable screening test for hypogammaglobulinemia in children less than 18 years of age.

Complete DPYD genotyping combined with dihydropyrimidine dehydrogenase phenotyping to prevent fluoropyrimidine toxicity: A retrospective study.

In April 2019, French authorities mandated dihydropyrimidine dehydrogenase (DPD) screening, specifically testing uracilemia, to mitigate the risk of toxicity associated with fluoropyrimidine-based chemotherapy. However, this subject is still of debate as there is no consensus on a standardized DPD deficiency screening test. We conducted a real-life retrospective study with the aim of assessing the impact of DPD screening on the occurrence of severe toxicity and exploring the potential benefits of complete genotyping using next-generation sequencing. All adult patients consecutively treated with 5-fluorouracil (5-FU) or its oral prodrug at six cancer centers between March 2018 and February 2019 were considered for inclusion. Dihydropyrimidine dehydrogenase deficiency screening included gene encoding DPD (DPYD) genotyping using complete genome sequencing and DPD phenotyping (uracilemia or dihydrouracilemia/uracilemia ratio) or both tests. Associations between each DPD screening method and (i) severe (grade ≥3) early toxicity and (ii) fluoropyrimidine dose reduction in the second chemotherapy cycle were evaluated using multivariable logistic regression analysis. Furthermore, we assessed the concordance between DPD genotype and phenotype using Cohen's kappa. A total of 551 patients were included. Most patients were tested for DPD deficiency (86%) including DPYD genotyping only (6%), DPD phenotyping only (8%), or both (72%). Complete DPD deficiency was not detected in the study population. Severe early toxicity events were observed in 73 patients (13%), with two patients (0.30%) presenting grade 5 toxicity. Despite the numerically higher toxicity rate in untested patients, the occurrence of severe toxicity was not significantly associated with the DPD screening method (p = 0.69). Concordance between the DPD genotype and phenotype was weak (Cohen's kappa of 0.14). Due to insufficient numbers, our study was not able to demonstrate any added value of DPYD genotyping using complete genome sequencing to prevent 5-FU toxicity. The optimal strategy for DPD screening before fluoropyrimidine-based chemotherapy requires further clinical evaluation.

Association of plasma uracil concentration with 5-FU pharmacokinetics, dose-limiting toxicity, and DPYD genotype in patients with gastrointestinal cancer.

751 Background: Plasma uracil concentration >16 ng/mL is used in some countries as a screening test for DPD deficiency. We conducted this exploratory study to evaluate the association of uracil concentration with 5-FU exposure, toxicity, and DPYD genotype in patients without known DPD deficiency. The main hypothesis was that lower uracil concentration would be predictive of subtherapeutic 5-FU drug exposure (5-FU AUC < 20 mg*h/L) in patients treated with infusional 5-FU. Methods: The study prospectively enrolled patients with GI cancer who were receiving initial chemotherapy with a regimen containing a 46-hour 5-FU infusion initially dosed at 2400 mg/m2. Study candidates were screened for the DPYD *2A, *13 and c.2846A>T gene variants; variant carriers were excluded. Participants gave pretreatment fasting morning blood samples for determination of plasma uracil concentration and DPYD whole-gene sequencing. Plasma samples were collected at hour 45 of 5-FU infusion in chemotherapy cycles 1 and 2 for determination of 5-FU exposure, measured as the 5-FU AUC. We estimated a linear mixed-effects model to test the association of uracil concentration with 5-FU AUC, adjusting for cycle, sex, creatinine, and 5-FU infusion dose (in mg/m2). Results: There were 29 evaluable participants with a median age of 64 years (range 41-81); nine (31%) were female. The most common cancer sites were colorectal (n = 15) and pancreatic (10). The median (IQR) plasma uracil concentration was 10.4 ng/mL (7.2, 12.6). Data for 5-FU AUC were available for 19 subjects in cycle 1 (median = 22.0 mg*h/L) and 27 subjects in cycle 2 (median = 19.3 mg*h/L). In the mixed-effects model, male sex was significantly associated with 5-FU AUC (beta = -12.0 [95% CI -17.2, -6.7], p <0.001) but plasma uracil concentration was not (beta = 0.35 [-0.16, 0.86], p=0.16). Holding other covariates constant, male sex was associated with a 29% decrease in 5-FU AUC (vs female) and plasma uracil concentration in the lowest quartile was associated with a 12% decrease in 5-FU AUC (vs highest quartile). Three of 29 participants (10%, all male) had uracil concentrations of >16 ng/mL, suggestive of DPD deficiency. The subject with the highest uracil concentration (23.1 ng/mL) had severe toxicity in cycle 1 (neutropenia, mucositis) requiring 5-FU dose reduction; whole gene sequencing identified a rare DPYD missense variant in this subject (c.2185G>A [p.A729T]). The other two subjects with uracil concentration >16 ng/mL did not have culprit DPYD gene variants and tolerated infusional 5-FU at 2400 mg/m2 without early dose-limiting toxicity. Conclusions: Our findings suggest that male sex is more important than low pretreatment plasma uracil concentration as a predictor of risk for subtherapeutic 5-FU drug exposure. This result is consistent with prior evidence that men experience less 5-FU associated toxicity than women.

Prevalence and molecular analysis of glucose-6-phosphate dehydrogenase deficiency in Chin State, Myanmar.

Glucose-6-phosphate dehydrogenase (G6PD) deficiency is caused by X-linked recessive disorderliness. It induces severe anemia when a patient with G6PD deficiency is exposed to oxidative stress that occurs with administration of an antimalarial drug, primaquine. The distribution of G6PD deficiency remains unknown while primaquine has been used for malaria treatment in Myanmar. This study aimed to investigate the prevalence of G6PD deficiency and its variants in Chin State, Myanmar. Among 322 participants, 18 (11 males and 7 females) demonstrated a G6PD deficiency. Orissa variant was dominant in the molecular analysis. This would be related to neighboring Indian and Bangladeshi population, in which Orissa variant was also reported as the main mutation type. The screening test for G6PD deficiency before primaquine treatment appears to be important in Myanmar.

Calculated Globulin Is Clinically Useful as a Screening Test for Antibody Deficiency in Turkish Adult Patients

Introduction: Heterogeneous clinical features of antibody deficiency (AD) may cause diagnostic delays. Calculated globulin (CG) (total protein minus albumin) has been proposed as a screening test to prevent morbidity due to diagnostic delays in AD. Our aim was to validate CG as a screening test for AD in Turkish adult patients by comparing its role with gamma globulin analysis in protein electrophoresis. Methods: Fifty serum samples were randomly collected for each level of CG from 15 to 25 g/L and tested for serum IgG, IgA, IgM levels and protein electrophoresis. Cut-off values predicting low IgG levels were calculated for electrophoretically determined gamma globulin and CG. Additionally, the data of 47 patients followed up in our clinic with a diagnosis of primary antibody deficiency (PAD) were retrospectively analyzed. Results: A total of 550 adult patients were included in the study. The CG value predicting patients with IgG <6 g/L as a screening test was determined as <20 g/L with 83.8% sensitivity and 74.9% specificity. The gamma globulin value which predicted patients with the same IgG value of 89.0% sensitivity and 89.4% specificity was determined as <7 g/L. In the retrospective analysis, 37 of 47 patients (78.7%) with PAD had a CG value of <20 g/L at the time of the diagnosis and all 13 patients (100%) whose gamma globulin values were measured at the time of the diagnosis had a gamma globulin value of <7 g/L. Conclusion: The determined CG cut-off value of <20 g/L can be used as a screening test in Turkish adult patients.

Maternal diabetes and the role of neonatal reticulocyte hemoglobin content as a biomarker of iron status in the perinatal period.

We aimed to evaluate the effects of maternal diabetes on neonatal iron status, measuring erythrocyte indices including hemoglobin, hematocrit, reticulocytes, mean corpuscular volume (MCV), percent (%) hypochromia, ferritin, and additionally mean reticulocyte hemoglobin content (MCHr) as an early marker of iron deficiency, and examine the association between neonatal MCHr, red cell indices, and ferritin. We conducted a hospital-based prospective cohort study in a tertiary neonatal unit of a University Hospital from 2018 to 2020. We enrolled 126 maternal-infant pairs of mothers whose pregnancy was associated with diabetes and 74 maternal-infant pairs from uncomplicated pregnancies. Erythrocyte indices were analyzed within the first twelve hours after birth. Erythrocyte parameters were compared between infants of the diabetes and the non-diabetic group. We examined the correlation of the neonatal MCHr with perinatal characteristics, including gestation, birth weight, maternal body mass index, the erythrocytic indices, maternal diabetes, maternal obesity, prematurity, small-for-gestational-age status, maternal preeclampsia, and maternal anemia. Finally, we evaluated the discordance between neonatal MCHr and neonatal ferritin. Infants of the diabetes group had a significantly lower MCHr (32.6 pg vs. 34.2 pg, p=0.003) compared with infants of uncomplicated pregnancies. Neonatal MCHr was significantly correlated with maternal hypochromia (r=-0.237, p=0.004) and neonatal MCV (r=0.674, p<0.001). Neonatal MCHr was significantly associated with maternal diabetes [standardized coefficients 0.21, 95% confidence interval (CI) 0.05-0.58, p=0.003) and maternal preeclampsia (standardized coefficients 0.17, 95% CI 0.02-0.92, p=0.019), after adjusting for maternal anemia, maternal obesity, prematurity, and small-for-gestational-age status. Those results were consistent also when analyzing maternal-infant pairs with pre-existing diabetes, and maternal-infant pairs with gestational diabetes. There was significant discordance between neonatal MCHr and neonatal ferritin (p=0.001). MCHr was significantly lower in infants of mothers whose pregnancy was associated with diabetes compared with infants of non-diabetic mothers and correlated with neonatal and maternal red cell indices of iron deficiency. Since there was significant discordance between neonatal MCHr and ferritin during the first postnatal day, it is possible that MCHr could be used as a screening test for iron deficiency, especially in infants.

Screening for Color Vision Deficiency in Health Science Students

Purpose: Various professions require employees to be able to detect different shades of color accurately when visually reading test results with colorimetric end points. To prepare health science students to better meet this requirement, a screening test for color vision deficiency (CVD) was administered to detect any major or minor errors as part of the undergraduate student experience. Method: Screening for color vision deficiency was administered using the Farnsworth D-15 Color Vision Test. Students completed the test and their results were scored and interpreted at the time of completion. Results: Students (n=85) from nine different health science programs completed the Farnsworth D-15 Color Vision Test. Ages of the participants ranged from 18-63 years with 70% of the participants in the age range of 18-23 years. Seventy-one percent of the participants identified as female, 28% identified as males, and 1% identified as non-binary. Two students (one female and one male) had a minor error resulting in a crossover within the test circle on the Gulden test score sheet. There were no major errors identified in the 85 participants. Conclusion: While two minor errors were detected in this population, this data is not consistent with other CVD studies. This most likely is due to screening predominately female students, and/or the amount of time allotted for each individual test to be completed. Future studies will include expanding the participant numbers, with a focus on testing more male participants, limiting the number of minutes to complete the screening process, and collecting additional demographic data.

Sustainability and clinical outcomes of routine screening for pathogenic DPYD gene variants prior to fluoropyrimidine (FP) chemotherapy for gastrointestinal (GI) cancer.

216 Background: Dihyropyrimidine dehydrogenase (DPD) deficiency is present in 3-5% of patients, and is associated with substantially increased risk of severe and/or fatal toxicity during standard-dose FP chemotherapy. Genotyping of pathogenic DPYD variants is a readily available screening test for DPD deficiency, and prospective studies show that dose-reduced FP chemotherapy can be used safely in heterozygous DPYD variant carriers. Methods: Following a sentinel toxicity event the GI medical oncology group at the Norris Cotton Cancer Center adopted a shared practice of routine screening for pathogenic DPYD gene variants prior to FP chemotherapy (5-FU or capecitabine). Screening procedures involved physicians, NP/PAs, nurses, pharmacists, and schedulers. Testing was completed at a send-out lab until late 2020, when an in-house test became available. The current test panel evaluates for 3 gene variants: c.1905+1G &gt; A (*2A), c.1679T &gt; G (*13), and c.2846A &gt; T. We report on the sustainability and clinical outcomes of DPYD gene variant screening. We identified all patients starting new FP-containing intravenous chemotherapy regimens (e.g., FOLFOX, CAPOX) for treatment of GI cancer at two sites (LEB &amp; STJ) between Jan. 2019 and May 2021. We used electronic medical records to evaluate for completion of DPYD genotyping, and we describe the prevalence and management of DPYD gene variant carriers. Results: We identified 333 patients starting FP-containing chemotherapy regimens during the study period, including 287 patients without prior history of FP chemotherapy. Screening with DPYD genotyping was completed in 228 of 287 eligible patients (79%). Screening rates increased from 34% in Q1 of 2019 to 90% in Jan-May 2021. Five GI oncology sub-specialists accounted for 89% of screen-eligible patients and 96% of completed tests, but 10 unique physicians ordered ≥1 test. Of 228 screened patients, six (2.6%) were heterozygous carriers of pathogenic DPYD gene variants (*2A [2 patients], *13 [1], and c.2846A &gt; T [3]). Variant carriers started FP chemotherapy with a 33-50% reduction. Two of six patients required further dose reduction due to FP-related toxicity (grade 4 neutropenia, grade 3 diarrhea). All evaluable variant carriers completed planned initial treatment. Implementation challenges included variable insurance coverage of DPYD genotyping, site-specific test ordering and reporting processes, and inconsistent turn-around time for send-out testing (resolved with on-site testing). Conclusions: Routine screening for pathogenic DPYD gene variants prior to FP chemotherapy is feasible and sustainable in the U.S. DPYD genotyping coupled with chemotherapy dose reductions for DPYD variant carriers facilitates safe and timely completion of planned chemotherapy treatments.

PathogenicDPYDVariants and Treatment-Related Mortality in Patients Receiving Fluoropyrimidine Chemotherapy: A Systematic Review and Meta-Analysis

Pathogenic variants of the DPYD gene are strongly associated with grade≥3 toxicity during fluoropyrimidine chemotherapy. We conducted a systematic review and meta-analysis to estimate the risk of treatment-related death associated with DPYD gene variants. We searched for reports published prior to September 17, 2020, that described patients receiving standard-dose fluoropyrimidine chemotherapy (5-fluorouracil or capecitabine) who had baseline testing for at least one of four pathogenic DPYD variants (c.1129-5923C>G [HapB3], c.1679T>G [*13], c.1905+1G>A [*2A], and c.2846A>T) and were assessed for toxicity. Two reviewers assessed studies for inclusion and extracted study-level data. The primary outcome was the relative risk of treatment-related mortality for DPYD variant carriers versus noncarriers; we performed data synthesis using a Mantel-Haenszel fixed effects model. Of the 2,923 references screened, 35 studies involving 13,929 patients were included. DPYD variants (heterozygous or homozygous) were identified in 566 patients (4.1%). There were 14 treatment-related deaths in 13,363 patients without identified DPYD variants (treatment-related mortality, 0.1%; 95% confidence interval [CI], 0.1-0.2) and 13 treatment-related deaths in 566 patients with any of the four DPYD variants (treatment-related mortality, 2.3%; 95% CI, 1.3%-3.9%). Carriers of pathogenic DPYD gene variants had a 25.6 times increased risk of treatment-related death (95% CI, 12.1-53.9; p < .001). After excluding carriers of the more common but less deleterious c.1129-5923C>G variant, carriers of c.1679T>G, c.1905+1G>A, and/or c.2846A>T had treatment-related mortality of 3.7%. Patients with pathogenic DPYD gene variants who receive standard-dose fluoropyrimidine chemotherapy have greatly increased risk for treatment-related death. The syndrome of dihydropyrimidine dehydrogenase (DPD) deficiency is an uncommon but well-described cause of severe toxicity related to fluoropyrimidine chemotherapy agents (5-fluorouracil and capecitabine). Patients with latent DPD deficiency can be identified preemptively with genotyping of the DPYD gene, or with measurement of the plasma uracil concentration. In this systematic review and meta-analysis, the authors study the rare outcome of treatment-related death after fluoropyrimidine chemotherapy. DPYD gene variants associated with DPD deficiency were linked to a 25.6 times increased risk of fluoropyrimidine-related mortality. These findings support the clinical utility of DPYD genotyping as a screening test for DPD deficiency.

Investigation of Circulating Plasma Micro RNAs As Biomarkers for Iron Status in Blood Donors

Introduction: Ferritin is the major iron storage protein in the body and serum ferritin is the most widely used screening test both for iron deficiency and iron overload conditions. However, as ferritin is a positive acute phase response protein its usefulness is limited in the presence of either acute or chronic inflammation or liver disease. A more specific biomarker of iron status could therefore be of considerable value. MicroRNAs (miRNAs) are small non-coding RNA fragments that repress target gene expression post-transcriptionally within cells. MiRNAs can also be detected outside cells in various body fluids and these circulating forms may be useful as biomarkers for various pathological conditions. Studies indicate that certain miRNAs may play an important role in cellular processes regulating iron homeostasis including, miRNA-320 in post-transcriptional control of Transferrin Receptor 1 (TfR1) gene expression, and miR-200b in regulation of ferritin expression. The aim of this study was to determine whether circulating miRNAs in plasma could serve as biomarkers for iron status in healthy blood donors.Methods: Study participants were selected from new/active blood donors based on their latest historic ferritin results in the blood bank laboratory information system. Participants were divided into two groups, a) high ferritin (>150 μg/l, HF) and b) low ferritin (<15 μg/l, LF) arms. The study was divided into two phases; i) a pilot study (20 participants) and ii) a confirmation study (50 participants). After obtaining informed consent EDTA blood samples were collected and frozen plasma aliquots prepared by a standard method to minimize pre-analytical variation. Ferritin was measured in all participants with an automated immunoassay (Ortho Vitros®). Hepcidin and sTFR plasma levels were measured by ELISA (DRG Instruments GmbH) in the pilot study group to confirm iron status. MiRNA isolation and analysis was performed by a miRNA profiling service (Exiqon A/S, Vedbæk, Denmark). In the pilot-study 179 miRNAs were analyzed using commercially available qPCR assays (Serum/Plasma Focus Panel, Exiqon A/S). In the confirmation study a custom-made Pick-&-Mix miRNA qPCR panel was ordered including 13 candidate miRNAs chosen from the pilot-study data. Standard biostatistical methods were utilized for data analysis.Results: Serum ferritin measured on average 13.8 ± 3.8 μg/l in the pilot study low ferritin (LF) arm, compared to 231 ± 58.8 μg/l in the high ferritin (HF) group (p<0.001). Hepcidin plasma levels were markedly lower in the low ferritin (LF) arm compared to the high ferritin group (HF) (2.3 ± 0.6 vs. 17.8 ± 3.1 ng/ml, p<0.001) whereas sTfR1 levels were higher in the LF group (1.9 ± 0.4 vs. 1.1 ± 0.2 µg/ml, p<0,001) confirming the difference in iron status between the two groups. Quality control of raw miRNA qPCR data indicated good technical performance of the profiling experiments. In the pilot-study 106 of 179 miRNAs in the panel gave signals in all 20 samples and on average 153 miRNAs were detected in each sample. On unsupervised analysis by hierarchical clustering and principal component analysis (PCA) the samples did not cluster according to iron status (figure 1). When comparing HF vs. LF groups 17 miRNAs were found to be differentially expressed when using a significance level of 0.05, but only one assay (hsa-miR-34a-5p) passed Benjamini-Hochberg correction for multiple testing. However, this particular miRNA frequently gave low/absent signals in both groups and was not considered further. To confirm the findings of the pilot study thirteen miRNAs were chosen for a confirmation-study based on technical performance and expression in the pilot study. Unsupervised analysis of 50 samples did not reveal any difference in expression for the 13 selected miRNAs between the LF and HF groups. Furthermore, when comparing the two groups no miRNA was found to be differentially expressed when using a p-value cutoff of 0.05.Conclusion: These data do not suggest that those circulating plasma miRNAs analyzed by qPCR panels in this study should be considered promising biomarkers for iron status in healthy individuals. However, in the future, sensitive and comprehensive detection methods such as next-generation sequencing might reveal novel circulating miRNAs that correlate with iron stores. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

First trimester screening for pre-eclampsia and intrauterine growth restriction using three-dimensional Doppler angiography (SPIRIT): protocol for a multicentre prospective study in nulliparous pregnant women

IntroductionPre-eclampsia (PE) and intrauterine growth restriction (IUGR) are two major pregnancy complications, related to chronic uteroplacental hypoperfusion. Nowadays, there is no screening or diagnostic test for uteroplacental vascularisation deficiency in...

Urinary ferritin; a potential noninvasive way to screen NICU patients for iron deficiency.

Building on our previous study, showing a correlation between ferritin in serum and urine, we conducted a feasibility evaluation, measuring urinary ferritin as a potential noninvasive screening test for iron deficiency among NICU patients. This was a prospective analysis of paired serum/urine ferritin levels. We defined iron-limited erythropoiesis by a RET-He <5th percentile lower reference interval (<28 pg). We obtained 49 paired serum/urine samples from neonates judged as at-risk for iron deficiency. Urine ferritin ("corrected" for urine creatinine and specific gravity) correlated with serum ferritin (correlation coefficient of log10-transformed values 0.44). A corrected urine ferritin <12 ng/mL had a sensitivity of 82% (95% CI, 67-93%) and a specificity of 100% (CI, 66-100%) for detecting iron-limited erythropoiesis, with a positive predictive value of 100% (CI, 89-100%). Measuring urinary ferritin in NICU patients is feasible. Since low values identify iron-limitation, this could become a useful noninvasive screen.

Geographical Accessibility to Glucose-6-Phosphate Dioxygenase Deficiency Point-of-Care Testing for Antenatal Care in Ghana.

Background: Glucose-6-Phosphate Dehydrogenase (G6PD) deficiency screening test is essential for malaria treatment, control, and elimination programs. G6PD deficient individuals are at high risk of severe hemolysis when given anti-malarial drugs such as primaquine, quinine, other sulphonamide-containing medicines, and chloroquine, which has recently been shown to be potent for the treatment of coronavirus disease (COVID-19). We evaluated the geographical accessibility to POC testing for G6PD deficiency in Ghana, a malaria-endemic country. Methods: We obtained the geographic information of 100 randomly sampled clinics previously included in a cross-sectional survey. We also obtained the geolocated data of all public hospitals providing G6PD deficiency testing services in the region. Using ArcGIS 10.5, we quantified geographical access to G6PD deficiency screening test and identified clinics as well as visualize locations with poor access for targeted improvement. The travel time was estimated using an assumed speed of 20 km per hour. Findings: Of the 100 clinics, 58% were Community-based Health Planning and Services facilities, and 42% were sub-district health centers. The majority (92%) were Ghana Health Service facilities, and the remaining 8% were Christian Health Association of Ghana facilities. Access to G6PD deficiency screening test was varied across the districts, and G6PD deficiency screening test was available in all eight public hospitals. This implies that the health facility-to-population ratio for G6PD deficiency testing service was approximately 1:159,210 (8/1,273,677) population. The spatial analysis quantified the current mean distance to a G6PD deficiency testing service from all locations in the region to be 34 ± 14 km, and travel time (68 ± 27 min). The estimated mean distance from a clinic to a district hospital for G6PD deficiency testing services was 15 ± 11 km, and travel time (46 ± 33 min). Conclusion: Access to POC testing for G6PD deficiency in Ghana was poor. Given the challenges associated with G6PD deficiency, it would be essential to improve access to G6PD deficiency POC testing to facilitate administration of sulphadoxine-pyrimethamine to pregnant women, full implementation of the malaria control program in Ghana, and treatment of COVID-19 patients with chloroquine in malaria-endemic countries. To enable the World Health Organization include appropriate G6PD POC diagnostic tests in its list of essential in-vitro diagnostics for use in resource-limited settings, we recommend a wider evaluation of available POC diagnostic tests for G6PD deficiency, particularly in malaria-endemic countries.

The modified G6PD deficiency screening test

Glucose-6-phosphate dehydrogenase (G6PD) deficiency, which involves the mutation of the G6PD gene on the X chromosome, is the most common enzyme defect in humans. G6PD produces nicotinamide adenine dinucleotide phosphate (NADPH) for red blood cell protection from oxidative stress. Oxidative stress-induced red blood cell hemolysis in G6PD-deficient patients leads to anemia, jaundice, acute renal failure, and kernicterus in newborns. Currently, the fluorescent spot test (FST) is widely used for G6PD deficiency screening. This test detects NADPH production under UV light. This study purposed to modify the G6PD deficiency screening test based on NADPH production and the ability to detect the results of the color change in the color of the reaction. Blood samples of 60 subjects were collected and screened for G6PD deficiency using FST method. All samples were tested with the modified G6PD screening test and the quantitative assay kit. The results from the modified test showed the color red for normal cases, whereas the color brown indicated G6PD deficiency cases. The cutoff value of G6PD activity detected by the quantitative method was less than 2.9 U/gHb for G6PD deficiency. The precision of this study was 100 %. The results of sensitivity, specificity, negative predictive value (NPV), and positive predictive value (PPV) of the modified test were 100 %, 96.7 %, 100 %, and 96.7 %, respectively. The modified G6PD screening test can be used for the screening of G6PD deficiency in newborns, for support through providing more information, and for self-protection from oxidative stress.

Ispitivanje i upoređivanje troškova G6PD enzimskog skrining testa za novorođenu decu u odnosu na troškove lečenja pacijenata sa favizmom hospitalizovanih usled hemolize

Objective. We compared the glucose-6-phosphate dehydrogenase (G6PD) deficiency screening costs in neonates and treatment costs for patients with favism who were referred to the government hospitals of Isfahan with acute hemolysis. Methods. In this retrospective study, 116 patients with favism were selected, and their clinical data and treatment costs were recorded in Iranian Rials (IRR) and United States Dollars (USD). The costs of neonatal G6PD screening tests were estimated based on quantitative and qualitative screening kits by referring to each hospital's laboratories. Approximately 84760 births occurred in Isfahan province from October 2014 to October 2015. Results. Based on our results, 73 patients (62.9%) were males, and 43 of them (37.1%) were females. The causes for hemolysis were the consumption of broad beans in 78.4%, infection in 19.8%, and drugs (erythromycin and sodium cephalothin) in 1.7% of patients. The majority of patients (46.6%) received a single blood transfusion, and 18.9% received two or more transfusions, while 34.5% had received no blood transfusions. The mean and standard deviation of patients' medical expenses were 7,472,005±7,329,847 IRR (276.74±271.47 USD). The total cost of treatment for patients was 869,162,324 IRR (32,191.19 USD). The screening cost for all newborns from October 2014 to October 2015 with qualitative kits was 754,364,000 IRR (27,939.40 USD) and with the quantitative kits was 960,330,800 IRR (35,567.80 USD). Conclusion. The costs of the qualitative screening test for G6PD deficiency are lower than the costs of treatments of favism patients in Isfahan.

A cross-sectional study on parental awareness for newborn screening and assessment of the burden of congenital hypothyroidism and glucose 6-phosphate dehydrogenase deficiency

Introduction: Newborn Screening (NBS) has been one of the most successful health programs and of most paramount importance worldwide but not so in India. Due to the lack of awareness and paucity of laboratory facilities, the disease burden in our population has not yet been established. Aim and Objective: The aim and objective of the study was to assess the burden of Congenital Hypothyroidism (CH) and Glucose 6-Phosphate Dehydrogenase (G6PD) deficiency in this area by NBS tests and to assess the impact of awareness sessions on the response rate of parents for NBS. Material and Methods: The screening was conducted in 474 babies of age 48 h up to 8 weeks. The dried blood spots collected were subjected to the following analytical protocol. Thyroid-stimulating hormone (TSH) level and G6PD enzyme activity were analyzed by immunofluorescence method-based neonatal kits. All babies with a positive screening test for G6PD deficiency and CH were asked for venous confirmatory testing after 7 days. Results: The efficiency of the program for all live birth babies delivered in the institute was 92% (n = 410/445). It was 82% in the first phase of the study period and 98.5% in the second phase. Repeated training of nursing professional reduced the sampling errors from 14.7% in the first phase to 6.1% in the second phase. A total of 11 samples reflected high TSH values, of which one baby confirmed for CH. Of the 24 babies who screened positive for G6PD, four were confirmed for the same. The prevalence for CH and G6PD deficiency was, respectively, 1 in 462 (2/1000) and 4 in 462 (8.7/1000). Conclusion: Development of expansion of NBS program in the state should be made mandatory for all newborns. The recommendations include awareness among parents, during antenatal and postnatal period and also to health professionals and provision for laboratory facilities for NBS testing at low cost.

Performance of the Access Bio/CareStart rapid diagnostic test for the detection of glucose-6-phosphate dehydrogenase deficiency: Asystematic review and meta-analysis.

To reduce the risk of drug-induced haemolysis, all patients should be tested for glucose-6-phosphate dehydrogenase (G6PD) deficiency (G6PDd) prior to prescribing primaquine (PQ)-based radical cure for the treatment of vivax malaria. This systematic review and individual patient meta-analysis assessed the utility of a qualitative lateral flow assay from Access Bio/CareStart (Somerset, NJ) (CareStart Screening test for G6PD deficiency) for the diagnosis of G6PDd compared to the gold standard spectrophotometry (International Prospective Register of Systematic Reviews [PROSPERO]: CRD42019110994). Articles published on PubMed between 1 January 2011 and 27 September 2019 were screened. Articles reporting performance of the standard CSG from venous or capillary blood samples collected prospectively and considering spectrophotometry as gold standard (using kits from Trinity Biotech PLC, Wicklow, Ireland) were included. Authors of articles fulfilling the inclusion criteria were contacted to contribute anonymized individual data. Minimal data requested were sex of the participant, CSG result, spectrophotometry result in U/gHb, and haemoglobin (Hb) reading. The adjusted male median (AMM) was calculated per site and defined as 100% G6PD activity. G6PDd was defined as an enzyme activity of less than 30%. Pooled estimates for sensitivity and specificity, unconditional negative predictive value (NPV), positive likelihood ratio (LR+), and negative likelihood ratio (LR-) were calculated comparing CSG results to spectrophotometry using a random-effects bivariate model. Of 11 eligible published articles, individual data were available from 8 studies, 6 from Southeast Asia, 1 from Africa, and 1 from the Americas. A total of 5,815 individual participant data (IPD) were available, of which 5,777 results (99.3%) were considered for analysis, including data from 3,095 (53.6%) females. Overall, the CSG had a pooled sensitivity of 0.96 (95% CI 0.90-0.99) and a specificity of 0.95 (95% CI 0.92-0.96). When the prevalence of G6PDd was varied from 5% to 30%, the unconditional NPV was 0.99 (95% CI 0.94-1.00), with an LR+ and an LR- of 18.23 (95% CI 13.04-25.48) and 0.05 (95% CI 0.02-0.12), respectively. Performance was significantly better in males compared to females (p = 0.027) but did not differ significantly between samples collected from capillary or venous blood (p = 0.547). Limitations of the study include the lack of wide geographical representation of the included data and that the CSG results were generated under research conditions, and therefore may not reflect performance in routine settings. The CSG performed well at the 30% threshold. Its high NPV suggests that the test is suitable to guide PQ treatment, and the high LR+ and low LR- render the test suitable to confirm and exclude G6PDd. Further operational studies are needed to confirm the utility of the test in remote endemic settings.

Reticulocyte hemoglobin content (CHr) untuk skrining defisiensi besi pada ibu hamil

Reticulocyte hemoglobin content (CHr) for iron deficiency screening in pregnant womenBackground: Anemia in pregnancy is a major public health problem in developing countries. In Indonesia, such anemia is generally caused by nutritional deficiencies, particularly iron deficiency. Iron deficiency screening is needed to prevent the development of iron-deficiency anemia in pregnant women that may affect the health of the mother and fetus.Objective: The study objective was to determine the diagnostic validity of reticulocyte hemoglobin content (CHr) for iron deficiency screening in pregnant women.Method: This research is analytic observational research with a cross-sectional design. The subjects were 73 pregnant women in Bantul district with various gestational ages. CHr was examined with an Advia 120 automatic hematology analyzer. Examination of ferritin as gold standard uses chemiluminescens method with Elecsys 1010 immunology analyzer. Table 2x2 is used to test several CHr values in the CHr range for women of reproductive age of 28.8 - 34.5 pg.Results: Based on ferritin &lt;15 μg / L as a gold standard, 13 pregnant women were suffered from iron deficiency while others were normal.Conclusion: CHr at 30.7 pg cut off has a sensitivity of 92.31%; negative predictive value (NPV) 96.2%; and likelihood ratio (LR) produce conclusive changes in post-test probability. In this cut-off, CHr can be used as an iron deficiency screening test in pregnant women.

SUN-256 Limited Usefulness of Single IGF-I Measurement as a Screening Test for Growth Hormone Deficiency in Children and Adolescents

Introduction: IGF-I measurement has been proposed as a screening tool for the diagnosis of Growth Hormone Deficiency (GHD). However, few studies evaluated the accuracy of IGF-I measurement as a screening test and the results have been controversial. Objective: To evaluate the value of a single IGF-I measurement as screening test in the diagnosis of GHD in short children and adolescents. Patients and Methods: 439 short children (median-IQR age 10.5 (7.1-12.8) y, 273 boys), were included in this retrospective analysis. The patient group consisted of 102 subjects (54 boys) with GHD (peak GH <7.0 μg/L after two provocative tests with Arginine, Insuline Tolerance Test, and Clonidine), 60 of whom had GHD of organic or genetic origin (OGHD) and 42 were idiopathic (IGHD). The 337 subjects with a GH peak ≥7 μg/L and no other recognizable cause for their shortness were considered as controls. All provocative tests were performed between 8.00 and 9.00 am after fasting overnight, without steroid priming. GH and IGF-I were measured by the same chemiluminescence assay in all samples (Immulite, Siemens). Receiver operating characteristic (ROC) analysis was used to evaluate the optimal IGF-I cut-offs and the diagnostic accuracy of IGF-I. The analysis was repeated including only 67 patients with severe GHD (SGHD, peak GH <5.0 μg/L after two provocative tests): 54 organic/genetic (SOGHD) and 18 idiopathic (SIGHD). Results: IGF-I SDS was significantly lower in patients than in controls (-2 (-2.9 - -1.7), -1 (-1.8 - -0.1), respectively, p<0.0001). IGF-I SDS ≥ -2 was found in 42% and 69% of OGHD and IGHD patients, respectively. The Area Under the Curve (AUC, measure of discriminative ability of the test) of IGF-I SDS was 0.73. The IGF-I SDS cut-off of -2 had 50% sensitivity (Se), 80% specificity (Sp), 2.4 likelihood ratio for positive test results (LR+), and 74% diagnostic efficiency (Ef). The best pair of values for Se and Sp was found at IGF-I SDS cut-off of -1.8 (B cut-off, Se 60%, Sp 75%, LR+ 2.3, Ef 71%). For OGHD, AUC was 0.82. For cut-off -2 SDS: Se 60%, Sp 80%, LR+ 3, Ef 80%. B cut-off was -1.9 SDS (Se 68%, Sp 78%, LR+ 3.1, EF 77%). For IGHD, AUC was 0.61. For cut-off -2 SDS: Se 36%, Sp 79%, LR+ 1.7, Ef 77%. B cut-off was -1.4 SDS (Se 52%, Sp 65%, LR+ 1.5, Ef 64%). The results were similar when only patients with SGHD were analysed. Among them, 42% had IGF-I SDS ≥ -2. IGF-I SDS ≥ -2 was found in 41% of SOGHD and in 67% of SIGHD. Conclusions: In our large cohort of short children, IGF-I measurement has shown suboptimal accuracy in discriminating GHD from non-GHD patients. The accuracy is better for patients with organic or genetic GHD. IGF-I values should be interpreted in combination with other clinical and biochemical parameters, and cannot be used alone as a screening tool.

Red Cell and Reticulocyte Parameters for the Detection of Iron Deficiency in Pregnancy.

Iron deficiency is a common complication of pregnancy and may lead to anemia as pregnancy progresses. Routine screening tests in pregnancy include hemoglobin levels, but in most centers not a serum ferritin. Advances in red cell and reticulocyte indices on automated blood counters have the potential to detect iron deficiency earlier, but pregnancy is associated with a rapid expansion of the red cell mass and parameters based on the entire erythrocyte population are less sensitive to changes. The objective of this study was to assess whether the Red Cell Size Factor and associated reticulocyte based indices can enable single-platform iron deficiency screening in pregnancy. Pregnant women had ferritin measured with blood counts and reticulocytes on a Beckman DxH800. The ability of the red cell size factor (RSF) and mean reticulocyte volume (MRV) to detect iron deficiency (ferritin < 10 µg/L) or early iron deficiency (ferritin < 30 µg/L) was assessed by comparing receiver operator characteristic curves and areas under the curve (AUC). RSF (AUC 0.80) and MRV (AUC 0.80) were both acceptable for detecting iron deficiency, but were not statistically superior to mean cell volume (MCV; AUC 0.77, p = 0.1). However, the optimal cut point for MCV was 86 fL, well above the accepted lower limit of normal. All parameters were poor at detecting early iron deficiency. Iron deficiency can be detected in pregnancy with red cell and reticulocyte parameters. While a low MCV is suboptimal as a screening test for iron deficiency, an MCV of 86 fL provides similar performance to the other red cell parameters studied.