New Born Screening (NBS) is the practice of testing every infant for dangerous or potentially fatal diseases that are else not apparent at birth. Beforehand discovery and prompt treatment can make the difference between healthy development or lifelong impairment and possible death. Utmost common New Born diseases in India are Prematurity, Respiratory Dysfunction, natural Deformations, Neonatal Infections & Haemolytic conditions of Newborn.[1] Congenital Hypothyroidism (CHT) is defined as thyroid hormone deficiency present at birth, which could be moreover due to absent or underdeveloped thyroid gland (Dysgenesis) or because of deficient hormone conflation in a well- developed thyroid gland (Dyshormonogenesis). CHT must be diagnosed instantly because detention in treatment can lead to unrecoverable neurological poverties. NBS styles have led to earlier opinion and treatment of CHT performing in bettered neurodevelopmental issues. The thyroid gland secretes the hormones T3 & T4 under the influence of TSH. The thyroid hormones play an essential part in energy metabolism, growth and neurodevelopment. Specifically, it acts on neuronal isolation, synapsis development, & myelination in antenatal & invigorated period, regulating CNS development.[2] G6PD insufficiency- Glucose-6-phosphate-dehydrogenase insufficiency is one of the commonest mortal enzymopathies, caused by inherited mutations of the X- linked gene G6PD. Being X-linked it is more common in males. G6PD insufficiency makes the red blood cells largely vulnerable to oxidative damage, and increased vulnerability to haemolysis. utmost people remain asymptomatic throughout their continuance; still, numerous may develop acute and occasionally veritably severe. Haemolytic Anaemia, when ‘touched off’ by ingestion of Fava sap, or by several medicines (for e.g. Primaquine, Rasburicase) or infrequently by infection. Acute Haemolytic Anaemia can be managed effectively handed it is instantly diagnosed. [3] Symptoms of Acute Haemolysis associated with G6PD Deficiency include Jaundice, Abdominal/ Back Pain, Fatigue, Haemoglobinuria, Haemolytic Anaemia. [4] This study is planned to screen the babies for these 2 diseases, that is, CHT and G6PD Deficiency & give a detailed description of the results attained. We hope this study can give meaningful information for unborn exploration and eventually contribute to timely and better clinical operation of newborn babies harbouring these diseases.

This study is a type of descriptive cross-sectional Study. It was done by Department of Biochemistry in collaboration with the Department of Paediatrics in a tertiary care centre in Malwa region of Punjab. All newborns born in the Labour Room/Operation Theatre of tertiary care hospital and all newborns admitted to the Paediatric Department of the tertiary care centre less than 14 days were included with proper informed consent. Incidence of these disorders was calculated based on the results obtained between the timeline of 4.5 months. Newborns who have received blood transfusion were excluded in the study. Fresh Venous Blood Samples of the newborns were collected. For thyroid profile estimation- plain vials were used and for G6PD estimation- EDTA vials were used.[5] Venous Sample were taken from Day 3 – Day 5 of life.[6] In newborns who were admitted on greater than 5th day and less than 14th day in the hospital their venous Samples were taken on the respective date of admission. Blood samples were processed in the Biochemistry laboratory for the following parameters: NAME OF THE PARAMETERS/ METHOD(S) USED/ INSTRUMENTS USED • T3, T4 –COMPETITIVE CHEMILUMINESCENCE IMMUNOASSAY [7] on instrument MAGLUMI SNIBE • TSH- SANDWICH CHEMILUMINESCENCE IMMUNOASSAY [8] on instrument MAGLUMI SNIBE • G6PD LEVELS (QUANTITATIVE)- KINETIC [9] on instrument LABSYSTEM DIAGNOSTICS CLINICAL CHEMISTRY ANALYSER [9] The ones who were screened positive for Congenital hypothyroidism were confirmed by a Confirmatory test by collecting their venous sample within 2 weeks.[10] The results obtained were assessed quantitatively for any possible deficiency (out of the 2 mentioned i.e. CHT and G6PD Deficiency) for each newborn. The parents were counselled regarding the purpose of the study, to find out CHT & G6PD deficiency and hence early management of the same. Proper confidentiality of reports was maintained with respect to each newborn enrolled. Statistical Analysis: Descriptive analysis of various parameters mentioned in the study was done and the percentage deficiency of the two disorders was calculated via Microsoft Excel and Epi info.

A total of 258 newborns were enrolled, their blood samples sent for quantitative estimation of the above-mentioned parameters and, the relevant data was collected from the mother or any other attendant for a timeline of 4.5 months. Following which, towards the end of 4.5 months, the entire data was compiled & the following results were found. The results were as follows: TABLE I: DISTRIBUTION OF NEONATES ON THE BASIS OF VARIOUS GESTATIONAL PARAMETERS GENDER MALE 124 48% FEMALE 134 52% GESTATIONAL AGE PRETERM 88 34% TERM 170 66% POSTTERM 0 0% BIRTH WEIGHT LOW 87 33.7% NORMAL 166 64.34% HIGH 5 1.93% MODE OF DELEIVERY NVD 132 51% LSCS 126 49% The screening was done randomly (prior number was not fixed number for males and females) and total of 258 newborns were screened and on analysis it showed that there were 48% males and 52% females. The gestational age was asked to look for any relation whether these disorders are more in preterm born babies as compare to term babies. But no such pattern was seen. Birth weight was also measured to see whether these babies have low or high birth weight. But maximum number of newborns were having normal birth weight which showed that these congenital disorders have no relation with birth weight. The mode of delivery also had no relation with the congenital disorders. TABLE II: MEAN ±SD OF VARIOUS THYROID PARAMETERS AND G6PD Parameters Mean ±SD T3 1.33 ± 0.467 T4 10.59 ± 4.11 TSH 3.37 ± 3.55 G6PD 14.33 ± 2.75 T3, T4, TSH and G6PD were analysed in all the newborns and their Mean ±SD was calculated. TABLE III: DISTRIBUTION OF NEONATES (NUMBER AND PERCENTAGE) HAVING G6PD DEFICIENCY AND CONGENITAL HYPOTHYROIDISM ON THE BASIS OF THEIR GENDER Diseases on screening Males Females Total G6PD Deficiency 2 - 2 (out of 258) Congenital hypothyroidism - 2 2 (out of 258) G6PD Deficiency % 0.7% - 0.7% Congenital Hypothyroidism % 0% 0.7% 0.7% Overall G6PD deficiency was came out to be 0.7% and it was seen only in males. The total percentage of congenital hypothyroidism was 0.7% and it was seen that it was seen in case of females and not in males. TABLE IV: COMPARISON OF NORMAL AND ABNORMAL VALUES OF THYROID AND G6PD CALCULATING THEIR t and p VALUES Parameters N (%) Mean ± SD t values and p values T4 Normal 256 (99.22%) 10.588±4.1214 0.5103, p=0.6103 Abnormal range 02 (0.77%) 12.080±3.281 T3 Normal 256 (99.22%) 1.33±0.466 1.4723, p=0.1422 Abnormal range 02 (0.77%) 1.820±0.367 TSH Normal 256 (99.22%) 3.180±2.73 12.9178, p<0.0001 Abnormal range 02 (0.77%) 28.60±8.20 G6PD Normal 258 (99.22%) 14.406±2.64 4.8567, p<0.0001 Abnormal range 02 (0.77%) 5.3 ± 0.14 The normal values of thyroid parameters (T3, T4 and TSH) of 256 neonates were compared with abnormal values of 2 neonates using unpaired t test and difference came out to be significant with TSH as p value was <0.0001 whereas it came out to be insignificant with T3 and T4. Similarly, for G6PD the 2 abnormal values (out of 258 neonates) were compared with 256 normal values and here also the difference came out to be significant as p value was less than 0.0001.

Invigorated webbing for common metabolic and inheritable diseases should be an integral part of neonatal care as early discovery and treatment can help intellectual and physical blights and life- changing ailments. The list of conditions for which webbing is carried out differs from country to country, based on the conditions and available coffers. Universal webbing for about 40 to 50 metabolic diseases is obligatory in US, Europe, and numerous other countries across the world. Though universal webbing is a cost ferocious exercise, the benefits far exceed the cost as it helps in reducing the mortality and morbidity of these conditions. [11] The conditions for which neonatal webbing has been proposed in Indian script include congenital hypothyroidism, Congenital adrenal hyperplasia (CAH) and glucose-6-phosphate dehydrogenase (G6PD) insufficiency [12- 19] This study enrolled 258 babies out of which there were 48 males and 52 females. The details regarding mode of delivery, birth weight of the baby and gravid age at the time of delivery were asked but showed no relation with the pattern of the congenital diseases in babies. According to this study, babies with natural hypothyroidism were 2 in 258, i.e. 0.7% and that of G6PD is 2 in 258 i.e. 0.7%. The data was not matching with the study by Verma J et al [20] who set up out the birth prevalence of 1 in 2,000 for CH, 1 in 2,500 for CAH, and 1 in 125 for G6PD insufficiency indicates the ultimate as the most current. Another study by Kumari B et al [21] presented the results that as aggregate of 492 babes were born from January 2020 to December 2020, of which 369 babies were screened for CAH, CH and G-6-PDD. Of 369 babes, one case had a raised position of TSH, six cases (all males) had a raised position of 17- OHP and no case was set up with G-6-PD insufficiency. The webbing tests done by Patel S et al [22] reported elevated TSH situations in 1.6 of total population and G6PD enzyme insufficiency in 2.6 of all babies enrolled. Confirmational tests revealed that 4[3.1/ 1000] babies were declared positive for CH and 8[6.2/ 1000] were came to be G6PD deficient. As compare to the below studies the prevalence of congenital hypothyroidism is more in over studied area. The reason could be attributed to this region is known to have iodine insufficiency in the soil and in the cultivated vegetables. Although the universal iodization program has introduced the iodised swab still the population is deficient in iodine and develop goitre, and other thyroid complaint The other reasons that are there for the increase in the cases of thyroid diseases are incognizance of complaint among some sections of the society and autoimmunity, which is the most common reason for thyroid complaint. Stress is another factor attributed for the increase in autoimmune affiliated cases of thyroid complaint. Inheritable predilection is also one of the reasons for thyroid complaint. Drawbacks in the study: 1. Sample size was a bit smaller. We can plan further study with a bigger sample size. 2. Screening for other diseases can be included.

India has a significant burden of G6PD deficiency and congenital hypothyroidism, yet awareness and screening coverage remains inadequate, leading to complications. The prevalence of G6PD deficiency varies across different ethnic groups, necessitating region-specific research to guide targeted screening and management strategies. With increasing cases detected through newborn screening, research is needed to explore environmental, genetic, and iodine-related factors contributing to this trend in India. Developing affordable and scalable screening programs is crucial to ensure accessibility in rural and underserved areas, reducing long-term healthcare costs. Strong research data can drive policy changes to integrate universal newborn screening into India’s national health programs, ensuring early diagnosis and treatment. More studies are needed to assess the long-term developmental outcomes of affected children and improve treatment protocols tailored to the Indian population. Investigating the role of dietary habits, pollution, and maternal health can help in developing preventive strategies for these conditions. Expanding research in these areas can significantly improve early detection, treatment outcomes, and overall child health in India. Implications of the study: Early identification of congenital hypothyroidism and G6PD deficiency can lead to timely interventions, preventing long-term complications such as developmental delays, mental retardation, or life-threatening conditions like haemolytic anaemia. By conducting the study and engaging the community, there will be increased awareness of congenital hypothyroidism and G6PD deficiency, empowering parents, and caregivers with knowledge about these conditions. Screening can help reduce the burden of preventable disabilities and improve overall quality of life for affected newborns. The newborns who are found to be deficient for G6PD are counselled to avoid the precipitating factors like ingestion of fava beans, or to avoid intake of drugs like primaquine or some antibiotics. Parents need to be educated on the diagnosis of congenital hypothyroidism (CH), the importance of early and adequate treatment that will prevent poor neurodevelopmental outcomes. Clear and adequate instructions on the L-T4 administration should be provided for caretakers.

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