Eye Single Tube Red Cell Research Articles

Prevalence of Sickle Cell Anemia In Eastern India

AbstractAbstract 4822 Background:Sickle cell disease is an inherited disorder of hemoglobin synthesis. This is due to replacement of Valine for Glutamic Acid in position six of the Beta globin chain of hemoglobin. This genetic alteration yields unstable RBC which lasts for 10–20 days. In stressful conditions the cells become sickle shaped and get lysed. There are about 20 million people with sickle cell disease in India. During January 2009- May2010 camps were held in various parts of West Bengal, Jharkhand, Chattisgarh. Along with various mutations of thalassemia, we also observed sickle cell anemia among them. This triggered our interest to study the spectrum of the sickle mutation co-inheritant with different mutations such as Homozygous Sickle Cell, Sickle Cell-Beta0 Thalassemia, Sickle Cell-Beta+ Thalassemia, Severe β+ thalassemia genes, Moderate β+ thalassemia genes, Mild β+ thalassemia genes Sickle cell-HbE Thalassaemia, Sickle cell-HPFH Thalassaemia, in said part of India. Since Indian patients with SS disease had higher hemoglobin, red cell counts and higher HbF levels and lower HbA2, MCHC, MCV, and reticulocyte counts, hence a high hemoglobin is a risk factor for painful crises and may also be a risk factor for avascular necrosis of the femoral head, proliferative sickle retinopathy, and acute chest syndrome. Methods:We have screened 332 individuals in eastern part of India during the period January 2009- May 2010. 3ml of peripheral blood was collected in EDTA vial from each individual. NESTROFT (Naked Eye Single Tube Red Cell Osmotic Fragility Test) was performed on spot. Then Complete Blood Count was done within 24 hours of collection. HPLC (High Performance Liquid Chromatography) was performed to identify the samples for confirmation. In our observation in case of sickle cell anaemia HbF (Fetal haemoglobin), Hb (haemoglobin), MCV (mean corpuscular volume) values ranges between 0–10 %, ≤7-10g/dl, 65–90fl respectively. ARMS (Amplification Refractory Mutation System) PCR (polymerase chain reaction) was done to confirm the mutation. Result:LocationTotal no. of IndividualNumber of carrier with mutationNumber of patient with mutationJharkand1032917Chattisgarh115279West Bengal114317 Conclusion:Of the total samples collected in the camps held at various places of Jharkand, Chattisgarh & West Bengal 87 of them was carriers of sickle cell anemia. There was 7 homozygous (SS), 14 sickle beta, 12 double heterozygous for HPFH (High Persistance of Fetal Hemoglobin) & sickle cell anemia. In conclusion, the manifestations of sickle cell disease are influenced by a variety of other genetic and environmental factors. The occurrence of the disease against different genetic and environmental backgrounds provides experimental models that contribute to understanding the variability in clinical and hematological expression of the disease. Disclosures:No relevant conflicts of interest to declare.

Haemoglobinopathies and glucose-6-phosphate dehydrogenase deficiency in a malaria endemic region of Manipur, northeast India

This study deals with the screening of haemoglobinopathies and glucose-6-phosphate dehydrogenase (G6PD) deficiency among three population groups of Manipur, namely 302 Pangan (Manipuri Muslim), 300 Bamon (Manipuri Brahmin), and 111 Kabui (tribal) individuals. Blood sam- ples were collected by finger prick method and were screened for abnormal haemoglobins by using the Naked Eye Single Tube Red Cell Osmotic Fragility Test. The male samples of Pangan (136), Bamon (127), and Kabui (51) were also screened for G6PD deficiency by the Fluorescent Spot Test. The alarmingly high frequencies of these two genetic disorders demand an immediate attention towards the general awareness for their prevention and management in Manipur and other neighbouring states.

Effort to Protect Totos, the Smallest Tribe in the World.

Abstract 5113 BackgroundToto is a primitive Indo-Bhutanese tribe residing in a small enclave called Totopara in the Jalpaiguri district of West Bengal, India. Toto culture and language is totally unique to the tribe, and is clearly distinguished from the neighboring Rajbongshis, Koch or the Bhutanese Sharchop tribes. Totos use to do consanguineous marriage and stay in the same locality even after marriage. Totos are becoming extinct since 1950. Total population of Totos are 1377 (smallest tribe of the world) and it is declining day by day. We noticed that their life span is very short (35-40 years). With a motive to investigate the reason behind it we started our work and got the answers of our queries. Almost 50% of them are carrier of beta Thalassemia, which is a genetic disease and are caused by the mutation of beta globin gene. The objectives of our study are,•To screen their whole population for Thalassemia (carrier / patient).•To increase the awareness level by arranging awareness program in that locality frequently through seminar, documentaries and discussions.•To make Totos understand about the ill fate of Thalassemia patients.•To control the birth of Thalassemic children, we are insisting them not to marry between two carriers.•Finally to confirm the mutation in globin gene by DNA analysis. Materials & MethodsAfter awareness program, 3-4 ml of blood sample was collected from each Toto along with their written consent. On the spot inexpensive NESTROF (Naked Eye Single Tube Red Cell Osmotic Fragility) test was done. After returning to institute we use to do CBC (Complete Blood Count) followed by HPLC (High Performance Liquid Chromatography). DNA was isolated from those samples detected as carrier or patients using the protocol standardized in our lab. Mutation was detected by ARMS (Amplification Refractory Mutation System)–PCR. ConclusionTotal individual already screened in the Totopara under the age limit 10- 35 years is 688. Number of Totos and Non-Totos are 628 and 60 respectively. From NESTROFT and CBC result it was revealed that different blood parameters e.g., MCV, MCH, RBC, RDW, Hb% etc of 71% Totos do not match with the normal value and found to be significantly altered. HPLC results show that 51% Totos are carrier of HbE and 20% of them are HbE homozygous. Whereas 97% non-Totos are found to be normal. DNA analysis by ARMS-PCR confirmed the result of HPLC. The percentage of HbE carrier and HbE homozygous is very high among Totos and this is the real cause of their early death. To protect this primitive tribe we have to restrict their marriage between two carriers. For Totos thalassemia carrier detection should be mandatory before marriage. DisclosuresNo relevant conflicts of interest to declare.

Prevalence of Thalassemia Among Rabhas; a Small Tribe in Eastern Part of India.

Abstract 5114 BACKGROUNDThalassemia syndromes are a heterogeneous anemia caused by mutation affecting globin chains of the hemoglobin molecule. It is estimated that the average life span of Rabhas (tribal population of North Bengal, Jalpaiguri) range from 42 – 45 years. This raised a concern that they are perishing due to some genetic disease. Hence Screening Camps were conducted to identify the reason. Rabha is a little known Scheduled Tribe community of West Bengal. Assam now renamed to Asom is a North-East India state of India with its capital at Dispur. The Rabha people are mainly found in the jungles of Jalpaiguri. West Bengal is a States and territories of India in eastern India. With Bangladesh, which lies on its eastern border the state forms the ethno-linguistic region of Bengal. Jalpaiguri is the largest district of North Bengal, covering an area 6,245 sq.km. It is situated between 26 16' and 27 0' North latitudes and 88 4' and 89 53' East longitudes and Cooch Behar district. Cooch Behar is a district of the state of West Bengal, India, as well as the name of the town which gives its name to the district. The whole area of Eastern and Western Dooars, may be termed as the cradle land of the Rabhas. The Rabhas belong to Indo-Mongoloid group of people and have similarities with other members of Bodo. Bodo may stand for: *Bod A city in Norway* An ethnic community in India: the Bodo people*The Bodo language spoken by them. group such as Garo (tribe). The Garos are a tribe in Meghalaya, India, and Mymensingh District, Bangladesh, who call themselves Achik. Kachari. The Kacharis are the most widely spread tribe in northeast India. They are said to be the earliest inhabitants of the Brahmaputra Valley. Mech (tribe), Koch. Our objectives were- Awareness among the tribal population through talks and documentaries and discussions with community leaders with demonstrations at village level. The goal of thalassemia screening is to identify the carrier status among Rabha populations, to control the birth of affected children thus eradicating thalassemia among them & to save one of the oldest tribe from being extinct. MATERIAL & METHODSAt first an Awareness Programme was held among the Rabhas & then with the written consent peripheral blood was collected for thalassemia screening test. The screening age lies between 10 – 35 years. Firstly, NESTROFT (Naked Eye Single Tube Red Cell Osmotic fragility Test) was performed for spot detection. This was followed by CBC (Complete Blood Count) & HPLC (High performance Liquid Chromatography) for confirmation.Molecular Analysis of every sample was done using ARMS PCR.All together 277 individuals were screened. Of which 119 (43%) were HbE carrier & 110 (40%) were HbE homozygous. Rest of them was normal. The carrier & homozygous status was confirmed by performing ARMS PCR. The sensitivity of NESTROFT in this case was 95 %. CONCLUSIONThus the percentage of HbE carrier & HbE homozygous is very high among the Rabhas. But one thing is to be noted that their % of haemoglobin is very high. This might be one of the reasons that they do not require blood transfusion during their life span. It is spreading like a rapid fire due to consanguineous marriage among them. This is one of the reasons for the early mortality. In our project of Department of Biotechnology, Govt. of India we'll complete carrier status detection of total Rabha population (11,000) within 3 years. DisclosuresNo relevant conflicts of interest to declare.

Is NESTROFT sufficient for mass screening for β-thalassaemia trait?

Prevention of beta-thalassaemia trait will, for the foreseeable future, hinge on effective screening strategies. Routine use of haematological data from automated cell counters may complement the results of the Naked Eye Single Tube Red cell Osmotic Fragility Test (NESTROFT), especially because of the high cost of a false-negative error. Our objective was to assess the potential additive value of routine haematological data in screening for beta-thalassaemia trait. Community survey of asymptomatic volunteers. Using the NESTROFT results, haematological data and haemoglobin A(2) concentration from 1435 young, asymptomatic Sindhi subjects recruited in a population-based survey, and statistical analysis by classification tree approach, we examined whether haematological parameters have discriminatory utility additional to that of NESTROFT in screening for beta-thalassaemia trait. We observed that in the derivation subset from which the classification tree was generated, there was only a marginal--albeit statistically significant--improvement in the screening performance of NESTROFT, whereas there was no such improvement attributable to the use of haematological parameters in a separate validation subset. Our results further substantiate the claim that the use of NESTROFT is highly indicated for screening for beta-thalassaemia trait in regions where the prevalence is high and the resources are constrained.

Prevalence of β-thalassaemia in subcastes of Indian Sindhis: Results from a two-phase survey

To estimate the prevalence of beta-thalassaemia in different subcastes of the Indian Sindhi population who, in general, have a high prevalence of this disease. A two-phase, community-based survey. Asymptomatic, Sindhi volunteers from Nagpur, central India, were recruited into the present study over a 7-year period. The first phase included the use of the Naked Eye Single Tube Red cell Osmotic Fragility Test (NESTROFT). Those positive for NESTROFT or those volunteering for haemoglobin A(2) (HbA(2)) quantification entered the second phase of the survey. Appropriate statistical methods for estimating prevalence from two-phase surveys were used. The prevalence of beta-thalassaemia carriers across the five major Sindhi subcastes varied substantially in the study population. Larkana Sindhis had the highest (17%) whereas Dadu Sindhis had the lowest (8%) frequency of the beta-thalassaemia allele. As a corollary, the projected incidence of beta-thalassaemia major in newborn babies greatly varied by the subcastes of the parents. Ethnic subgroups within populations known to commonly carry the beta-thalassaemia gene provide further information that is useful from epidemiological and public health perspectives.

Bias-corrected diagnostic performance of the naked eye single tube red cell osmotic fragility test (NESTROFT): An effective screening tool for β-thalassemia

It is being increasingly recognized that a majority of the countries in the thalassemia-belt need a cost-effective screening program as the first step towards control of thalassemia. Although the naked eye single tube red cell osmotic fragility test (NESTROFT) has been considered to be a very effective screening tool for β-thalassemia trait, assessment of its diagnostic performance has been affected with the reference test- and verification-bias. Here, we set out to provide estimates of sensitivity and specificity of NESTROFT corrected for these potential biases. We conducted a cross-sectional diagnostic test evaluation study using data from 1563 subjects from Central India with a high prevalence of β-thalassemia. We used latent class modelling after ensuring its validity to account for the reference test bias and global sensitivity analysis to control the verification bias. We also compared the results of latent class modelling with those of five discriminant indexes. We observed that across a range of cut-offs for the mean corpuscular volume (MCV) and the hemoglobin A2 (HbA2) concentration the average sensitivity and specificity of NESTROFT obtained from latent class modelling was 99.8 and 83.7%, respectively. These estimates were comparable to those characterizing the diagnostic performance of HbA2, which is considered by many as the reference test to detect β-thalassemia. After correction for the verification bias these estimates were 93.4 and 97.2%, respectively. Combined with the inexpensive and quick disposition of NESTROFT, these results strongly support its candidature as a screening tool—especially in the resource-poor and high-prevalence settings.