Gene prevalence in abnormal haemoglobin divergent and blood groups in Uttarakhand

Context: Abnormal haemoglobinopathis are generally observed deviant disarray happen due to genetic inequality within the alpha and beta chains amino acids sequences alters. Aims and objectives: a total of 933 cases were included in the study for detection of Thalassemic cases. Out of these positive thalassemic cases were further analysed for gene frequency. Materials and Methods : Starting with complete Blood Cell counts which has led to suspected cases for further analysis by haemoglobin electrophoresis by HPLC (high performance liquid chromatography) for confirmation of Thalassemic cases. Finally gene frequency detection by ARMS (Amplification Refractory Mutation System) by using PCR (Polymerase Chain Reaction). Isolation of DNA from whole blood by commercially available kit (QIA amp DNA blood midi kit 100 samples). Result: A sum total of 933 subjects, aged between 01-30 yrs were studied and it depicts 4.1% prevalence. β -thalassemia trait was screened as highest and S-D disease as lowest. The frequencies with respect to ABO is shown as B>0>AB>A. The amplicons which were analysed after gel electrophoresis were screened for ARMS, PCR, which IVS1-5 (G-C) Maccounts for almost 50.0% and lowest is Fr 8/9 (+G) M is 12.1%. Conclusion: As such patients requires repeated blood transfusion so availability of maximum type of affected Blood group is the time of need for availability to with blood banks. Further genetic studies will definitely help in effective for further pharmaceutical companies. © 2020 Published by Innovative Publication. This is an open access article under the CC BY-NC license (https://creativecommons.org/licenses/by-nc/4.0/)


Introduction
The beginning elaborations started in 1930 on Tunisians people accumulating date of relationship of blood groups (ABO) system with altered haemoglobinopatheis. [1][2][3][4] This cumulative work with multiple speculations along with altered haemoglobinopatheis was initial of its kind. 4 Further many observers has developed more precise method for the detection of β -thalassemia mutations in South East Asia, based on PCR generated restriction sites which are very synonymous to β -thalassmia mutations and later on practised in ruling out pathological mutations in mitochondrial DNA. 5 These modifications are observed at positions IVS1-5, IVS1-1 of β -globin gene. 6-8

Diagnostic criterion
By performing complete Blood Cell Counts; By HPLC (high performance liquid chromatography) for the detection and confirmation of Thalassemic cases; by (Bio-Rad D-10) haemoglobin testing system. By analysis by ARMS (Amplification Refractory Mutation System); PCR (Polymerase Chain Reaction) for gene frequency distribution. Isolation of DNA from whole blood which was preserved in anticoagulant (EDTA) by using the already commercially available kit (QIA amp DNA blood midi kit 100 samples).

Results
That the total number of 933 cases included in the study population we found blood group B (313) was observed to be more frequent and least is O (148). Among Rh positive male, blood group B (192), and in female, blood group A (87) was most prevalent blood group (Table 1).
While observing the frequency of ABO Rh blood group in abnormal haemoglobin variants, which were accounted as 40 cases. Within these blood group B positive (42.5%) is observed maximally and blood group A positive (9.7%) is least ( Table 2).
The frequency of mutation detected in abnormal haemoglobin variants are 36, out of total 40 cases included in the study. The mutation IVS 1-5(G→C)M(50.0%) is maximum and frequently observed and the least in Fr 8/9 (+G) M (12.1%) sharing with codon 41/42 N (TC TT). While 04 cases were uncertain in which study were insufficient for analysis (Table 3).
If we segregate the abnormal haemoglobionopathies according to gender then we observe the males (27) are more sufferer then females (13) ( Table 4).
As far as we study abnormal haemoglobinopathies distribution according to age & sex wise then we observe that more cases in males were observed during 0 to 10 yrs and then subsequently decreases but in females the least between the age 11 to 15 yrs, lesser between 0 to 5yrs but maximum cases were observed between the age 6 to 10 yrs (Table 5).

Discussion
The sequel of abnormal haemoglobinopathies are moreover autosomal recessive disorders and are genetically inherited through one or both parents who might be the carrier or suffering in another presentable form with the disease. 9,10 An aggregate of 933 patients were analysed for abnormal haemoglobin variants, ABO & rhesus blood groups ranging from 01 to 30 years. Out of the total 933 subjects 629 were males and 304 were females. Table 1 depicts the dispersal of the blood group (ABO) & Rhesus (D) between study subjects. Blood group B was observed as the majority frequent 313 while blood group O was least frequent 148. Here we observe the same frequency in earlier studies. 11,12 In Rh (D) blood typing 667 was Rh positive and 266 was Rh negative. Amongst Rh positive male blood group B (192) was observed as the most prevalent blood group proceeding ahead by blood group A (130), AB (73), O (66).
This shows that such mutations in thalassemic patients are more prevalent in males then females. Table 5 depicts that males shows more features and incidence during the age between 0 to 5 and 6 to 10 yearsparallarly, but decreases on increasing age. This may be because of less life survival rate after the age of 10 years. In comparison to males, females thalassmemic cases are more observed during the age between 6 to 10 years, then 0 to 5 years. This may be because of negligence and laid back attitude towards females in male dominance society.  A  130  34  87  37  217  71  288  B  192  36  58  27  250  63  313  AB  73  44  48  19  121  63  184  O  66  54  13  15  79  69  148  Total  461  168  206  98  667 266 933

Conclusion
These varied type of abnormal haemoglobinopathies with particular to thalassamia are the greatest factor of genetic mutational anamoly, which has eventually lead to wide spread public health disorder, therefore because clinical importance after birth. Elaborate study of varied haemoglobinopathies and their screening will definitely be a center stone while observed their occurrences with every region of the state along with the data shoring with regional research center -precise by formulating a data of blood groups in relation to abnormal haemoglobinopathies may furnish details about the available of human blood during emergencies, and also enlightens the possibility of future burden of disease.

Source of Funding
None.

Conflict of Interest
None.