THE USE OF HPLC AS A TOOL FOR NEONATAL CORD BLOOD SCREENING OF HAEMOGLOBINOPATHY - A VALIDATION STUDY

Background: Newborn cord blood screening identifies infants with underlying haemoglobinopathies before they develop the characteristic symptoms or sequelae. Aims: This study was performed to validate the interpretation high-performance chromatography (HPLC) along with complete blood count (CBC) results as a tool for universal neonatal screening of hemoglobin disorders in Oman. Methods: HPLC and CBC data on subjects who participated in the National Neonatal screening program at birth were obtained from archival records. The results recorded at birth were compared with a second study performed on the same subjects, after approval from the local medical research and ethics committee.Results: Only 290 subjects from amongst the original cohort of 3740 newborns could be recalled between April 2010 to March 2011, to repeat HPLC and CBC, as well as perform confirmatory DNA studies, wherever necessary. All these subjects had been documented to show an initial abnormal result. 31 cases who had no HbA at birth on HPLC were confirmed as either homozygous β-thalassaemia major (n=5 subjects) or homozygous sickle cell anemia (n=26 subjects) by appropriate DNA analysis. Additionally, amongst 151 subjects, 72 subjects were studied in the initial study by Hb Bart’s quantitation using aalpha thalassaemia short program at birth. In this cohort, 42 subjects with Hb Bart’s >1% at birth could be confirmed as having either deletional or non-deletional thalassaemia by GAP PCR studies. No case of HbH was detected in this cohort. Further, carrier status for structural hemoglobin variants (HbS, HbC, HbD, HbE) (n=67) and beta thalassaemia allele with low HbA at birth (n=29 out of 41) were confirmed by relevant molecular studies.Conclusions: The study validated the earlier observation by 100% concordance with results of CBC and HPLC. Presence of Hb Bart’s at birth does not always mean the presence of alpha thalassemia, as subjects with Hb Bart’s was below 1% by quantitation, were shown to be normal by molecular studies. Key Words: Neonatal, screening, HPLC validation, haemoglobinopathy, sickle cell disease, thalassaemia

Download Full-text

Fatal Acute Splenic Sequestration at 4 Months of Age

PEDIATRICS ◽

10.1542/peds.73.4.507 ◽

1984 ◽

Vol 73 (4) ◽

pp. 507-508

Author(s):

Juan N. Walterspiel ◽

Joe C. Rutledge ◽

Bryan L. Bartlett

Keyword(s):

Cord Blood ◽

Sickle Cell ◽

Sickle Cell Anemia ◽

Screening Program ◽

Splenic Sequestration ◽

Blood Screening

A patient with homozygous sickle cell anemia is the youngest known to have died from acute splenic sequestration crisis. A cord blood screening program might have prevented this infant's death.

Download Full-text

Screening Cord Bloods for Detection of Sickle Cell Disease in Jamaica

Clinical Chemistry ◽

10.1093/clinchem/20.6.666 ◽

1974 ◽

Vol 20 (6) ◽

pp. 666-669 ◽

Cited By ~ 87

Author(s):

Beryl E Serjeant ◽

Miriam Forbes ◽

Leslie L Williams ◽

Graham R Serjeant

Keyword(s):

Sickle Cell Disease ◽

Cord Blood ◽

Sickle Cell ◽

Screening Program ◽

Blood Collection ◽

Cell Disease ◽

Agar Gel ◽

Large Populations ◽

Cord Blood Collection ◽

Blood Screening

Abstract A cord-blood screening program, designed primarily for detecting sickle cell disease, has been in operation for seven months (8000 samples) at a large maternity unit in Kingston, Jamaica. We describe techniques of cord-blood collection and electrophoretic investigation on both cellulose acetate and agar gel. These methods appear to give rapid, valid results at minimal expense and are well adapted to screening large populations.

Download Full-text

The Neonatal Screening Program in Brazil, Focus on Sickle Cell Disease (SCD)

International Journal of Neonatal Screening ◽

10.3390/ijns5010011 ◽

2019 ◽

Vol 5 (1) ◽

pp. 11 ◽

Cited By ~ 1

Author(s):

Ana Silva-Pinto ◽

Maria Alencar de Queiroz ◽

Paula Antoniazzo Zamaro ◽

Miranete Arruda ◽

Helena Pimentel dos Santos

Keyword(s):

Sickle Cell Disease ◽

Sickle Cell ◽

Health Care Professionals ◽

Neonatal Screening ◽

Screening Program ◽

Cell Disease ◽

Ministry Of Health ◽

Basic Care ◽

Main Challenge ◽

Neonatal Screening Program

Since 2001, the Brazilian Ministry of Health has been coordinating a National Neonatal Screening Program (NNSP) that now covers all the 26 states and the Federal District of the Brazilian Republic and targets six diseases including sickle cell disease (SCD) and other hemoglobinopathies. In 2005, the program coverage reached 80% of the total live births. Since then, it has oscillated between 80% and 84% globally with disparities from one state to another (>95% in São Paulo State). The Ministry of Health has also published several Guidelines for clinical follow-up and treatment for the diseases comprised by the neonatal screening program. The main challenge was, and still is, to organize the public health network (SUS), from diagnosis and basic care to reference centers in order to provide comprehensive care for patients diagnosed by neonatal screening, especially for SCD patients. Considerable gains have already been achieved, including the implementation of a network within SUS and the addition of scientific and technological progress to treatment protocols. The goals for the care of SCD patients are the intensification of information provided to health care professionals and patients, measures to prevent complications, and care and health promotion, considering these patients in a global and integrated way, to reduce mortality and enhance their quality of life.

Download Full-text

Identification of the hot-spot areas for sickle cell disease using cord blood screening at a district hospital: an Indian perspective

Journal of Community Genetics ◽

10.1007/s12687-015-0223-7 ◽

2015 ◽

Vol 6 (4) ◽

pp. 383-387 ◽

Cited By ~ 5

Author(s):

Sujata Dixit ◽

Pushpansu Sahu ◽

Shantanu Kumar Kar ◽

Sapna Negi

Keyword(s):

Sickle Cell Disease ◽

Cord Blood ◽

Sickle Cell ◽

District Hospital ◽

Hot Spot ◽

Cell Disease ◽

Indian Perspective ◽

Blood Screening

Download Full-text

Cord blood screening for hemoglobin abnormalities by thin layer isoelectric focusing

Blood ◽

10.1182/blood.v56.6.1068.bloodjournal5661068 ◽

1980 ◽

Vol 56 (6) ◽

pp. 1068-1071 ◽

Cited By ~ 1

Author(s):

F Galacteros ◽

K Kleman ◽

J Caburi-Martin ◽

Y Beuzard ◽

J Rosa ◽

...

Keyword(s):

Thin Layer ◽

Cord Blood ◽

Sickle Cell ◽

Isoelectric Focusing ◽

Sickle Cell Trait ◽

Cell Disease ◽

Abnormal Hemoglobin ◽

Excellent Method ◽

Hemoglobin Variants ◽

Blood Screening

Hemoglobin variants can be successfully identified in cord blood samples. The methods most commonly used include cellulose acetate (CAC) and citrate agar (CAG) electrophoresis. Recently thin layer isoelectric focusing (TLIF) has been shown to be an excellent method for identifying hemoglobin variants. To determine the applicability of TLIF for cord blood screening, we compared the results of 835 samples obtained by TLIF with that obtained by CAC, CAG, and the combination of both CAC and CAG. In 100 of these samples we detected an abnormal hemoglobin pattern using TLIF. In contrast, we detected only 80 abnormal samples by CAC, 70 by CAG, and 80 by using the combination of CAC and CAG. Due to the increased resolution provided by TLIF, we correctly diagnosed two sickle cell trait samples by TLIF that were incorrectly suspected to be homozygous for sickle cell disease by CAC and CAG. We identified 41 samples containing Bart's hemoglobin by TLIF in contrast to only 21 using CAC and 14 using CAG. The time and cost of TLIF was comparable to that using the combination of both methods. We, therefore, conclude that TLIF is the method of choice for cord blood screening.

Download Full-text

Cord blood screening for hemoglobin abnormalities by thin layer isoelectric focusing

Blood ◽

10.1182/blood.v56.6.1068.1068 ◽

1980 ◽

Vol 56 (6) ◽

pp. 1068-1071 ◽

Cited By ~ 37

Author(s):

F Galacteros ◽

K Kleman ◽

J Caburi-Martin ◽

Y Beuzard ◽

J Rosa ◽

...

Keyword(s):

Thin Layer ◽

Cord Blood ◽

Sickle Cell ◽

Isoelectric Focusing ◽

Sickle Cell Trait ◽

Cell Disease ◽

Abnormal Hemoglobin ◽

Excellent Method ◽

Hemoglobin Variants ◽

Blood Screening

Abstract Hemoglobin variants can be successfully identified in cord blood samples. The methods most commonly used include cellulose acetate (CAC) and citrate agar (CAG) electrophoresis. Recently thin layer isoelectric focusing (TLIF) has been shown to be an excellent method for identifying hemoglobin variants. To determine the applicability of TLIF for cord blood screening, we compared the results of 835 samples obtained by TLIF with that obtained by CAC, CAG, and the combination of both CAC and CAG. In 100 of these samples we detected an abnormal hemoglobin pattern using TLIF. In contrast, we detected only 80 abnormal samples by CAC, 70 by CAG, and 80 by using the combination of CAC and CAG. Due to the increased resolution provided by TLIF, we correctly diagnosed two sickle cell trait samples by TLIF that were incorrectly suspected to be homozygous for sickle cell disease by CAC and CAG. We identified 41 samples containing Bart's hemoglobin by TLIF in contrast to only 21 using CAC and 14 using CAG. The time and cost of TLIF was comparable to that using the combination of both methods. We, therefore, conclude that TLIF is the method of choice for cord blood screening.

Download Full-text

Alpha-gene deletions in black newborn infants with Hb Bart's

Blood ◽

10.1182/blood.v56.5.931.bloodjournal565931 ◽

1980 ◽

Vol 56 (5) ◽

pp. 931-933

Author(s):

K Ohene-Frempong ◽

E Rappaport ◽

J Atwater ◽

E Schwartz ◽

S Surrey

Keyword(s):

Cord Blood ◽

Dna Analysis ◽

Newborn Infants ◽

Older Children ◽

Gene Deletions ◽

Alpha Thalassemia ◽

Definitive Method ◽

Alpha Gene ◽

Hb Bart's

The presence of increased Hb Bart's (gamma 4) in cord blood is believed to be an indication of alpha-thalassemia. We have used restriction endonuclease nalyses of DNA to compare the number of alpha-genes with the percentage of Hb Bart's in 6 older children who had Hb Bart's at birth and 17 newborns. Four children with > 2% Hb Bart's had Eco R1 alpha-gene patterns and hematologic data consistent with the presence of two alpha-genes, one per chromosome. Of the remaining 19 children, all of whom had < 2% Hb Bart's, 8 had 3, while 11 had 4 alpha-genes. Three infants with Hb Bart's between 1% or less Hb Bart's. infants with 3 alpha-genes may therefore have elevated or normal levels of Hb Bart's at birth. DNA analysis is the definitive method for the determination of heterozygous alpha-thalassemia syndromes in newborns.

Download Full-text

Alpha-gene deletions in black newborn infants with Hb Bart's

Blood ◽

10.1182/blood.v56.5.931.931 ◽

1980 ◽

Vol 56 (5) ◽

pp. 931-933 ◽

Cited By ~ 12

Author(s):

K Ohene-Frempong ◽

E Rappaport ◽

J Atwater ◽

E Schwartz ◽

S Surrey

Keyword(s):

Cord Blood ◽

Dna Analysis ◽

Newborn Infants ◽

Older Children ◽

Gene Deletions ◽

Alpha Thalassemia ◽

Definitive Method ◽

Alpha Gene ◽

Hb Bart's

Abstract The presence of increased Hb Bart's (gamma 4) in cord blood is believed to be an indication of alpha-thalassemia. We have used restriction endonuclease nalyses of DNA to compare the number of alpha-genes with the percentage of Hb Bart's in 6 older children who had Hb Bart's at birth and 17 newborns. Four children with > 2% Hb Bart's had Eco R1 alpha-gene patterns and hematologic data consistent with the presence of two alpha-genes, one per chromosome. Of the remaining 19 children, all of whom had < 2% Hb Bart's, 8 had 3, while 11 had 4 alpha-genes. Three infants with Hb Bart's between 1% or less Hb Bart's. infants with 3 alpha-genes may therefore have elevated or normal levels of Hb Bart's at birth. DNA analysis is the definitive method for the determination of heterozygous alpha-thalassemia syndromes in newborns.

Download Full-text