Premarital Screening Research Articles

Knowledge and Attitudes towards Thalassemia among Public University Students in Bangladesh

Background: Thalassemia is the most common congenital single-gene condition globally, characterized by a lack of or reduced synthesis of either the α- or β-globin chains and passed down from parents to offspring. This study aimed to determine the level of knowledge of thalassemia among Bangladeshi students from universities and their opinion on prevention. Methods: A cross-sectional descriptive online survey was conducted on public university students in Bangladesh using a structured questionnaire between June and November 2020, completed by students online. Demographic characteristics, knowledge, and attitude of the participants were assayed with 10 questions on each topic. The data were analyzed using Python. Descriptive statistics methods such as frequencies and percentages were used to present data. Result: A total of 681 students participated in the online survey. The average age of the respondents in this study was 21.97 years, with a standard deviation of 2.9. Most of the students, 611(89.72%), had heard about Thalassemia, yet only 248 (36.42%) students had good knowledge of Thalassemia. The knowledge level of students had no significant relationship with sex (p-value = 0.0819), marital status (p-value = 0.2281), and year of study (p-value = 0.4619), but had a considerable difference with the field of study (0.0042). However, 478 (78.36%) participants showed a positive attitude toward "Premarital Screening" to prevent Thalassemia. Conclusions: Long-term and target-based preventive initiatives are recommended for university students and the general population of Bangladesh, where the rate of consanguineous marriage is high. These initiatives would provide crucial information and increase awareness of premarital screening and genetic counseling to prevent thalassemia.

Premarital Screening for Thalassemia Among Young Adults: A Study in Zabol, Sistan and Baluchistan Province of Iran

Introduction: There is a high prevalence of hemoglobinopathies, including thalassemia, in Sistan and Baluchistan province of Iran. It is important to appropriately screen young adults to control the birth of new cases of thalassemia. In many less-equipped centers, this practice only relies on cell blood count. This study investigated the hematological parameters of young adults referring to the Central Laboratory of Sistan, Zabol, for premarital screening for thalassemia. Methods: This was a descriptive cross-sectional study conducted in Zabol, Sistan and Baluchistan province of Iran from August 2020 to August 2021. Complete blood count (CBC) parameters of 2926 young adults were collected using a Sysmex KX-21 device and were compared with available reference intervals according to the Clinical and Laboratory Standard Institute. Results: The means of women’s and men’s age were 38.84±16.32 and 36.64±14.14 years, respectively. The average values for each parameter in the studied population were red blood cell (RBC) (4.78±0.55), hemoglobin (13.19±2.46 g/dL), hematocrit (40.11±3.57%), mean corpuscular volume (MCV) (85.16±28.33), neutrophil (52.12±9.98), lymphocyte (37.84±9.03), mixed leukocytes (10.00±3.51), platelets (244.02±62.32). The mean values of hemoglobin (14.48±5.17 vs. 12.69±1.39 g/dL), hematocrit (43.01±3.89 vs. 38.97±3.45%), serum ferritin (115.4±74.3 vs. 69.20±12.36 g/dL), total iron-binding capacity (316.84±1.38 vs. 320.95±41.01, P=0.01), and serum iron (85.05±28.51 vs. 76.25±28.22, P=0.02) were significantly lower in women than in men. Out of 2,926 people who entered this study, 195 (6.66%) were suspected of having thalassemia because of their low MCV (71.61±7.33 fL) and mean corpuscular hemoglobin (MCH) (23.67±2.66 pg). Conclusion: The relatively low hemoglobin in females referring for premarital screening for thalassemia awakens healthcare experts to carefully examine these cases for the possibility of thalassemia minor after excluding other common possible reasons such as iron deficiency anemia using iron studies and hemoglobin electrophoresis.

PREVALENCE OF SICKLE-CELL DISEASE IN SAUDI ARABIA: A SYSTEMATIC REVIEW

Background: A series of hemoglobinopathies known collectively as sickle cell disease (SCD) contain abnormalities in the gene encoding the beta component of haemoglobin. There are other subcategories that fall under the SCD umbrella, including sickle cell disease (SCD), haemoglobin SC disease (HbSC), and haemoglobin sickle-beta thalassemia (beta-thalassemia positive or beta-thalassemia negative). The prevalence of SCD varies greatly across Saudi Arabia, with the Eastern province having the greatest frequency and the southern regions having the second-highest prevalence. The reported sickle-cell prevalence ranges from 2% to 27%, and in some regions, up to 2.6% of people will have SCD.
 Objectives: The study aims to summarize current evidences regarding Prevalence of Sickle-Cell Disease in Saudi Arabia.
 Methods: For article selection, the PubMed database and EBSCO Information Services were used. All relevant articles relevant with our topic and other articles were used in our review. Other articles that were not related to this field were excluded. The data was extracted in a specific format that was reviewed by the group members.
 Conclusion: Although the prevenance of sickle cell anemia is relatively high due to multiple reasons such as consanguinity, the prevalence of genetic diseases in Saudi Arabia may be significantly lowered during the following decades as a result of premarital screening there. Also, acute chest syndrome in SCD patients is relatively infrequent in Saudi Arabia's Eastern Province, it nonetheless has a major impact on morbidity and death. If patients with African haplotypes are compared, it has a low prevalence and recurrence.

Study of Prevalence of Thalassemia trait among couples undergoing premarital examination in Al-Hamdaniya

Background and objectives:
 Thalassemia is the most common single genetic disorder. Being a preventable disease, the “premarital screening program” was established. The objective of the study was to determine the prevalence of β-thalassemia trait among couples attending a center for premarital screening in Al-Hamdaniya district.
 Material and methods:
 A cross-sectional study was carried out in Al-Hamdaniya district (located in the north of Iraq) during December 2019 through July 2020 that included 600 healthy couples screened for hemoglobinopathies. Complete blood count (CBC) and HB electrophoresis were done.
 Results:
 The prevalence of abnormal Hb F and Hb A2 was 4.8% and 5.8% respectively but the differences were not significant between males and females. No significant association was detected between the degree of consanguinity with the prevalence of Hb F and the prevalence of Hb A2. And there was no significant association between family history of haemolytic disease and the prevalence of Hb F and prevalence of Hb A2
 Conclusion:
 The prevalence of thalassemia trait in Al-Hamdaniya, like other cities in the north of Iraq is high, so premarital screening program is beneficial for identification and prevention of risky marriages.

Identification and functional characterization of a novel homozygous intronic variant in the fumarylacetoacetate hydrolase gene in a Chinese patient with tyrosinemia type 1

BackgroundHereditary tyrosinemia type 1 (HT1; OMIM# 276700) is a genetic metabolism disorder caused by disease-causing variants in the fumarylacetoacetate hydrolase (FAH) gene encoding the last enzyme of the tyrosine catabolic pathway. Herein, we describe the clinical features and genetic characteristics of HT1 in a five years and seven months old Chinese patient.MethodsAfter clinical diagnosis of the proband with HT1, genetic testing was performed by Sanger sequencing of the FAH gene in all family members. Functional analysis of the disease-causing variant was performed by cDNA sequencing to understand the effect of the variant on FAH transcript. To further predict the variant effect, we used Human Splicing Finder (HSF) and PyMol in silico analysis.ResultsWe identified a novel previously undescribed intronic variant in the FAH gene (c.914-1G>A). It was detected in a child who was homozygous for the variant and had the clinical presentation of HT1. cDNA sequencing showed that this splice-junction variant affected the transcription of FAH by formation of two different transcripts. Our observations and laboratory experiments were in line with in silico methods.ConclusionsOur study provides new insight into the HT1 variant spectrum and a better understanding of this disease in the Chinese population. This will be useful for molecular diagnosis in our country in cases where premarital screening, prenatal diagnosis and preimplantation genetic diagnosis are planned.

Characterization of the Molecular Spectrum of a-Thalassemia Mutations in the Western Province of Saudi Arabia and Recommendation for Premarital Screening

Characterization of the Molecular Spectrum of a-Thalassemia Mutations in the Western Province of Saudi Arabia and Recommendation for Premarital Screening

Awareness of Sickle Cell Disease Among Adolescents and Young Adults

Background: Sickle hemoglobinopathy is the most common genetic disorder identified through the state mandated newborn screening program, occurring in 1:2,647 births and may pose an enormous health burden. While sickle cell disease is associated with significant morbidity, sickle cell trait is a benign condition. Due to this reason, knowledge of their sickle cell trait for an individual may be lacking while the condition may have been identified at birth. There were 2904 cases of sickle cell traits identified through the NJ newborn screening program in 2020. While other countries have programs to increase awareness of sickle hemoglobinopathy status to include premarital genetic screening, the implication of sickle cell trait on procreation and family planning are not highly emphasized in the US. Assessing awareness of sickle cell disease among adolescents is important given high rates of teenage pregnancies among this population. Through this study, we set out to assess awareness of sickle cell disease among adolescents. Methods: We performed a cross sectional study of 180 adolescents who sought medical care at one of the outpatient clinics or emergency room at Children's Hospital of New Jersey in Newark, NJ and administered an anonymous questionnaire to volunteers 12-21 years of age from January 2020-June 2022. The study was approved by our local IRB and an informed consent and/or assent was obtained from each enrolled subject. Questionnaires included a variety of questions including demographics and several questions to assess their knowledge of sickle cell disease. Questionnaires that were incomplete were not included in analysis. Results: Of the 180 adolescents, most were teenagers ages 14-18 years old (72%), majority identified as females (59%) and 92% were African Americans or Hispanic. More than 30% of subjects' parents were high school graduates, 25% were college graduates and the remainder had some other lower level of education. Out of 180,136 children (76%) had heard of sickle cell disease and only 78 children (44%) reported that they knew about sickle cell disease. However, when asked questions regarding sickle cell disease, only 12% were able to correctly identify that it is a genetic disease that may cause pain crises. One hundred and eighteen patients (66%) were unaware of their sickle cell status. Of those with sexual partners, 53% did not think that knowing their partner's sickle cell status would influence their choice and yet, 59% wouldn't consider having a child with someone who has sickle cell disease. Only 5 individuals (2.78%) were able to correctly identify the inheritance pattern of sickle cell disease. Of the 4 individuals with children, only 1 (25%) reported being aware of their child's sickle cell status. Ninety-two percent were interested in learning about sickle cell disease, and the majority identified their doctor, internet and school as primary forms of obtaining health information. Conclusion: Sickle cell disease is associated with significant health burden. While we cannot prevent sickle cell disease, as health care providers, we should make every effort to educate youth about this important condition. One of the ways is to review newborn screening hemoglobinopathy results during adolescent well visits, especially for those with sickle cell trait. Appropriate education and genetic counseling must be provided to these young individuals especially since many may engage in sexual activity at a young age.

Knowledge, Perception, and Attitude of Future Couples towards Premarital Screening

Knowledge, Perception, and Attitude of Future Couples towards Premarital Screening

The scenario of knowledge, attitude and practice of the Bangladeshi population towards thalassemia prevention: A nationwide study.

Thalassemia is one of the most common life-threatening yet preventable congenital hemoglobin disorders especially in South Asian regions like Bangladesh. It has become a rising public health concern for Bangladesh as 6-12% of the population are carriers and many of them are unaware of it. The purpose of the study is to inspect the knowledge and attitude towards thalassemia among the general people of Bangladesh. A cross-sectional survey was conducted in eight administrative regions of Bangladesh between January and October of 2020. A structured questionnaire was designed to collect information about thalassemia and socio-demographic characteristics. Multivariate logistic regression models were used to identify factors associated with knowledge of thalassemia. A p-value <0.05 was considered significant. Of the 1,248 participants, only 47.4% had heard of thalassemia. Half of the participants who heard about the disease had no idea that thalassemia was not a transfusion transmitted disease. Only 49.8% of participant correctly identified consanguineous marriages as an important risk factor. Majority of them knew that marriage between two carriers can lead to a child with thalassemia major. About 72.5% knew that blood tests are a diagnosis method to determine thalassemia. Among the socio-demographic variables, the level of education of the respondents was identified as an independent predictor for knowledge (p<0.05) on thalassemia. For example, graduate (aOR: 24.88; 95% CI: 6.238-99.232) or post-graduate (aOR: 33.18; 95% CI: 7.864-140.001) participants were more aware of thalassemia than non-graduates. However, about 68.2% of the participants showed a positive attitude towards premarital screening of themselves or their family members and 85.3% were willing to donate blood to thalassemia patients. The study shows that there is a need to disseminate the information on thalassemia since the knowledge gap is huge among people. These findings will strengthen the implementation of thalassemia major awareness through educational programs, health counseling, premarital screening and campaigning.

Community awareness of hereditary blood diseases and the benefit of premarital screening program in Saudi Arabia

Community awareness of hereditary blood diseases and the benefit of premarital screening program in Saudi Arabia

PROFIL KESEHATAN CALON PENGANTIN WANITA DI KANTOR URUSAN AGAMA KALIWATES JEMBER

The Premarital Check-up Program is one of the programs designed by the government to improve the quality of human resources for newborns, examination is used as a preventive, more precisely in premarital or before marriage in the hope of preparing for a healthy pregnancy in an effort to reduce AKI and AKB. The purpose of this activity is premarital check-up screening for prospective brides. This study uses direct observation and examination of prospective brides. The examinations carried out include physical examination of height, weight, LILA, blood pressure, laboratory examination of hemoglobin levels, glucose levels. The population in this study is the bride. The results of direct examination of the prospective bride, there are still prospective brides whose nutritional status is calculated from the count BMI is still in the poor category (21.3%) and LILA status is still in the poor category (17%), so education is needed about premarital check-up and preparation for being a healthy mother.

First and Common Clinical Presentation of Sickle Cell Disease in Children

Background: sickle cell disease (SCD) is the most common inherited blood disease worldwide with the vast majority of cases occurring in sub-Saharan Africa. In Sudan, sickle cell anemia is one of the commonest chronic hemolytic anemias with high mortality and morbidity. Objective: the objective of this study is to determine the first and common presentation of sickle cell in children to assist early diagnosis, prevent complications and decrease morbidity and mortality of sickle cell disease in children. Methodology and Results: the study represented a combined descriptive study (hospital-based) and analytic study (cross-sectional observational) at Mohamed El-Amin Hamed Pediatric Emergency Hospital for Children in Omdurman, Khartoum State where 50 children diagnosed with sickle cell anemia, for the first time, aged 4 months -17 years were included during period of six months (January – June 2016). The commonest first presentation of SCD was sickle cell crises in 46%, features of infection in 18% and the rest 36% presented with combined symptoms. 62% of children presented with pallor as a feature of anemia, 32% with jaundice, 14% with fatigability, 14% with shortness of breathing and 6% presented with other symptoms related to anemia. Conclusion and Recommendations: It is clear that features of vaso-occlusive crises were the first presenting clinical features of sickle cell disease followed by features of infections and anemia. We believe that starting premarital screening, neonatal screening, expand and support SCD clinics and stablishing social support networks will help in decreasing morbidity and mortality from SCD.

Mutation screening of the CYP1B1 gene reveals thirteen novel disease-causing variants in consanguineous Pakistani families causing primary congenital glaucoma.

BackgroundPrimary congenital glaucoma (PCG) is a heterogeneous rare recessively inherited disorder prevalent in regions with high consanguinity. Disease phenotype is associated with increased intra ocular pressure and is a major cause of childhood blindness. Sequence variations in Cytochrome P450 1B1 (CYP1B1) gene are a major cause of PCG. Current study was conducted to screen CYP1B1 gene in highly consanguineous PCG affected families from Pakistani population consistent with the autosomal recessive pattern of PCG inheritance.MethodsFor this study, patients and controls (clinically unaffected individuals of each family) from 25 consanguineous families belonging to Punjab, Baluchistan and Khyber Pakhtunkhwa, Pakistan were recruited through ophthalmologists. DNA was isolated from collected blood samples. Genetic screening of CYP1B1 gene was done for all enrolled families. In-silico analysis was performed to identify and predict the potential disease-causing variations.ResultsPathogenicity screening revealed sequence variants segregating with disease phenotype in homozygous or compound heterozygous form in eleven out of 25 analyzed families. We identified a total of sixteen disease causing variants among which five frameshift i.e., c.629dup (p.Gly211Argfs*13), c.287dup (p.Leu97Alafs*127), c.662dup (p.Arg222Profs*2), c.758_759insA (p.Val254Glyfs*73) and c.789dup (p.Leu264Alafs*63), two silent c.1314G>A, c.771T>G and six missense variations c.457C>G (p.Arg153Gly), c.516C>A (p.Ser172Arg), c.722T>A (p.Val241Glu), c.740T>A (p.Leu247Gln), c.1263T>A (p.Phe421Leu), and c.724G>C (p.Asp242His) are previously un reported. However two frameshift c.868dup (p.Arg290Profs*37), c.247del (p.Asp83Thrfs*12) and one missense variant c.732G>A (p.Met244Ile), is previously reported. Furthermore, six polymorphisms c.1347T>C, c.2244_2245insT, c.355G>T, c.1294G>C, c.1358A>G and c.142C>G were also identified. In the intronic region, a novel silent polymorphism i.e., g.35710_35711insT was found in homozygous state. All the newly detected disease-causing variants were negative in 96 ethnically matched controls.ConclusionAmong twenty-five screened families, eight families (PCG50, 52–54, 58, 59, 63 and 67) were segregating disease causing variants in recessive manner. Two families (PCG049 and PCG062) had compound heterozygosity. Our data confirms genetic heterogeneity of PCG in Pakistani population however we did not find molecular variants segregating with PCG in fifteen families in coding exons and intron-exon boundaries of CYP1B1 gene. Genetic counseling was provided to families to refrain from practicing consanguinity and perform premarital screening as a PCG control measure in upcoming generations.

Knowledge and Attitude toward Hemoglobinopathies in Premarital Screening Program among the General Population in the Western Region of Saudi Arabia

The National Premarital Screening Program, which includes sickle cell disease and thalassemia, was made mandatory in 2004 by the Kingdom of Saudi Arabia (KSA), and the earlier studies have shown a poor knowledge and negative attitude toward this program in the different study groups. This study was conducted to assess the knowledge and attitudes toward premarital screening (PMS) in a randomly selected national sample of the Saudi population, 18 years and above. This was a cross-sectional study conducted in the Saudi population in the western region between July and December 2021. Valid and reliable questionnaire and data were collected from 893 participants aged ≥18 years. The χ2 test was used to ascertain if there is an association between categorical variables. Multivariate logistic regression was used to determine factors predicting satisfactory knowledge. All 893 study participants had heard about PMS with 625 (70.0%), 244 (27.3%) and 24 (2.7%) having satisfactory, fair and poor knowledge, respectively. Participants aged 26–35 years (p =0 .038), females (p < 0.001), those with higher education (p = 0.003) and employed (p = 0.004), had a better knowledge compared to other groups. Most of the participants had a positive attitude toward PMS. There is a changing trend in the knowledge and attitude toward PMS with a greater number of people wanting to go for PMS. There is also an improvement in the number of participants opting out of marriage in case of incompatibility with their future partner. However, the health education programs need to be improved regarding the hemoglobinopathies.

Evaluation of the validity of the pre-marriage mean corpuscular volume value as a predictive test for b-thalassemia carrier status.

The national mandatory premarital screening test is based on mean corpuscular volume (MCV) > 80 fL value for the detection of β-thalassemia to provide acceptance for marriage. The objective of this study is to assess the efficacy of MCV as a screening test for β-thalassemia trait in the present population. This study was conducted on 418 blood samples collected from adult individuals. The diagnosis of β-thalassemia carrier was given to those having HbA2 values equal to or above 3.5%. The diagnostic reliability of different RBC indices and formulas in discriminating cases of β-thalassemia trait were evaluated. Finally, a new index called "Momani" was determined based on MCV, RDW and RBC count. β-thalassemia trait was identified in 10% of the cases. The measured MCV value was significantly lower in β-thalassemia carrier group compared to non-carrier group (p = <0.001). MCV value and RBC count showed a higher diagnostic reliability than other RBC indices. We found that MCV ≤ 74.45 fL is more suitable cutoff value of MCV with 86.2% specificity, 71.4% sensitivity, 36.6% positive predictive value, and 96.4% negative predictive value. Finally, our index "Momani" was found to be useful in predicting carrier and paralleled the performance of Sirdah, Mentzer, and Ehsani indices. MCV<80 is a useful but not a perfect cutoff point for the screening of β-thalassemia carriers from noncarriers. The diagnostic accuracy of MCV can be improved by selecting a new cutoff value. Moreover, "Momani" index shows good discrimination ability in diagnosing β-thalassemia carrier in our population.

Co-Inheritance of Heterozygous β0-Thalassemia with Single Functional α-Globin Gene: Challenges of Carrier Detection in Pre-Marital Screening Program for Thalassemia

This is a report of a couple with abnormal hematological indices who were investigated for α &amp; β-thalassemia mutations. Based on CBC and capillary hemoglobin electrophoresis results, the male and female subjects were β &amp; α-thalassemia carriers, respectively. Multiplex-Gap-PCR and Sanger sequencing techniques were used for the identification of mutations on α and β-globin genes. The DNA test showed the presence of c.315 + 1 G &gt; A mutation on β-globin gene of male subject while the female case had – MED double gene deletion and c.427T &gt; C mutation on α-globin and, interestingly, she was also a carrier for c.315 + 1 G &gt; A mutation on β-globin gene. Cases with the coinheritance of heterozygous β0-thalassemia with one functional α-globin gene have normal HbA2 levels that may lead to their being misdiagnosed as β-thalassemia carriers, especially in premarital screening programs for thalassemia. Therefore, β-globin gene sequencing is recommended in cases with normal Hb electrophoresis and reduced hematological indices in premarital screening programs for thalassemia, especially in regions with a high frequency of β-globin mutations, in order to identify all the β-thalassemia carriers.

P-097: KNOWLEDGE AND EXPERIENCES OF PARENTS WITH CHILDREN AFFECTED BY SICKLE CELL DISEASE FOLLOWED AT CHUK: A QUALITATIVE STUDY

Purpose: Parental knowledge about SCD has a direct effect on reducing mortality and complications related to Sickle cell disease. Families of children affected by SCD face several hardships caused by raising a child afflicted with this disease. However, parental knowledge about SCD and challenges faced by families of children with SCD have not been assessed in Rwanda. The aim of this study was to explore the knowledge on sickle cell disease among parents of children affected by SCD and to identify their experiences while caring for affected children. Materials and methods: A qualitative method using thematic analysis was used. Data collection was done using semi-structured interviews with thirteen parents from one referral hospital with Pediatric hematology clinic in Rwanda. Results: Thirteen caregivers including eleven mothers and two fathers participated in the study. Two broad themes were identified: knowledge and understanding of SCD, and experiences and hardships. The majority of parents had some knowledge of SCD but most of them had no knowledge of inheritance pattern of SCD. Prior to the diagnosis being made, all parents had never heard of SCD and they had misconceptions about the condition of their children. Parents face several challenges including persistent psychosocial distress and practical hardships caused by SCD. This study also revealed coping strategies used by parents. Conclusion: This study showed that parents had some knowledge of SCD with gaps of inheritance pattern. Families of children affected by SCD face several burdens including nutrition for the affected child, poverty and persistent emotional distress. Raising awareness of SCD among the public, availing premarital screening program for high risk people and providing social support for affected families would help to mitigate the impact of this disease. The authors do not declare any conflict of interest

Return on Investment from the Prevention of Spinal Muscular Atrophy in Kuwait

Background: Spinal muscular atrophy (SMA) is a rare genetic disease with a devastating nature and associated with significant clinical and economic burden. Fortunately, the availability of genetic screening and prevention technologies made preventive strategy possible. The aim of this study is to assess the return on investment from a prevention program that includes premarital genetic screening, preimplantation genetic diagnosis, and in vitro fertilization pertaining to SMA, spanning five-year period from the financial perspective of the Ministry of Health (MOH) in Kuwait. Methods: A stochastic financial model was developed following a return on investment (ROI) approach spanning five-year period. Results: An average ROI estimated to be 11% with cost saving amounted to 466,159 KWD (1,524,339 USD). The financial burden if the prevention program was not implemented is 16,441,539 KWD (53,763,832 USD). Conclusion: This study is one of the very few in the region to estimate the benefits of implementing a prevention program pertaining to SMA from the financial perspective of the MOH in Kuwait.

Cell-free fetal DNA as a non-invasive method using pyrosequencing in detecting beta-globin gene mutation: A pilot study from area with limited facilities in Indonesia.

BackgroundThalassemia is a monogenic, autosomal recessive, inherited disorder of the red blood cells caused by mutations or deletions in the globin gene. Approximately 6–10% of the Indonesian population carries the β-globin gene mutation; however, premarital screening is rarely conducted, and antenatal screening is optional. We explored the use of cell-free fetal DNA (cffDNA) as a potential non-invasive method of detecting the fetal β-globin gene mutation prenatally in pregnant women.Materials and methodsPregnant mothers (n = 10), who were known carriers of thalassemia and who had a history of having borne a baby with thalassemia major, and their carrier husbands (n = 4) were recruited after providing consent. EDTA blood was drawn, and maternal DNA, including cffDNA, and paternal DNA were isolated. Maternal contamination tests were conducted using the variable number tandem repeat test for ApoB and D1S80 loci. Allele quantification was performed by pyrosequencing. Known mutations from the bio-archived DNA of patients with thalassemia major (n = 16) were run alongside as a control.ResultsIn total, 7 out of 10 cffDNA successfully passed the maternal contamination test. The results of the allele quantification showed that six fetuses were predictive carriers of IVS1nt5 and one was predictive normal, in line with the allele quantification for the bio-archived DNA from patients with thalassemia major. The minimum threshold percentage for mutant A allele at cd26 was 32%, mutant T allele at IVS1nt1 was 23%, and mutant C allele at IVS1nt5 was 39%.ConclusionTaking cffDNA from the mother’s blood proved useful as a non-invasive means of detecting the β-globin gene mutation using pyrosequencing allele quantification. This non-invasive method is of great interest for prenatal diagnosis in settings with limited facilities, as it minimizes the risk of abortion. Further study of other mutations of the β-globin gene is needed.

The spectrum of Alpha and Beta Thalassemia Mutations: A 10-year Population-based Study of the Premarital Health Screening Program in West of Iran

Background: In various cancers, Ganoderic Acid A (GAA), an active triterpenoid derived from Ganoderma Background: Thalassemia refers to a category of inherited disorders resulting from defects in synthesizing one or several chains of hemoglobin (Hb). The present study aimed to determine the frequency of alpha and beta-thalassemia mutations in Kurdistan province, Iran.&#x0D; Materials and Methods: In this retrospective cross-sectional study, the laboratory data of 340 patients with thalassemia (170 females and 170 males), who were candidates for genetic testing in Kurdistan province, were examined over ten years (2006-2016). The participants were Kurd couples selected from the premarital health screening program.&#x0D; Results: In this demographic study, 20 beta mutations and nine alpha mutations were identified. Among the beta-thalassemia mutations, intervening sequence or intron No. 2- first nucleotide change as splice site mutation (IVS-II-1) (26.1%), codons 8/9 (14.8%), and intervening sequence or intron No. 1- first nucleotide (IVS-I-1) (12.2%) change as splice site mutation), had the highest frequency rates, respectively, constituting 53% of the beta mutations. In addition, α3.7(82.7%), -α4.2(8.3%), and --MED (Mediterranean deletion) (3.75%) were the most frequent alpha mutations, which constituted more than 90% of the alpha mutations.&#x0D; Conclusion: According to the results, the most frequent mutations in the HBB gene are IVS-II-1, Codons 8/9, and IVS-I-1, and in the HBA gene are α3.7, -α4.2, and –MED in Kurdistan province. In addition, the role of race and ethnicity as significant, influential factors in thalassemia was observable in the findings. The obtained results also indicated the communication pattern between the studied region's populations. Identifying common thalassemia mutations in an area could greatly benefit the early detection of thalassemia carriers in genetic laboratories and enhance thalassemia prevention programs.