md-medicaldata

Authors

Ivana Kavečan^1,2, Milan Obrenović², Tatjana Redžek Mudrinić^1,2, Nataša Nikolić³, Jadranka Maksimovic<sup>4

1</sup>Univerzitet u Novom Sadu, Medicinski fakultet, Novi Sad, Srbija
²Institut za zdravstvenu zaštitu dece i omladine Vojvodine, Novi Sad, Srbija
³Institut za javno zdravlje Vojvodine, Novi Sad, Srbija
⁴Univerzitet u Beogradu, Medicinski fakultet, Institut za epidemiologiju, Beograd, Srbija

 

UDK: 616-056.7:575 (497.113)"2021/2024"

The paper was received / Rad primljen: 14.02.2025.

Accepted / Rad prihvaćen: 31.03.2025.

 

Correspondence to:

Prof. dr Ivana Kavečan
Medicinski fakultet, Univerzitet u Novom Sadu
Institut za zdravstvenu zaštitu dece i omladine Vojvodine
Hajduk Veljkova 3
21000 Novi Sad
Srbija
tel. +381 63 580 552
e-mail: ivana.kavecan@mf.uns.ac.rs

 

 

Sažetak

 

 

Uvod: Implementacija molekularno genetičkih tehnika je značajan događaj za genetički centar i predstavlja sinhronizovane procese i niz komplementarnih aktivnosti radi uvođenja novih tehnologije u rutinski rad, kao i savladavanje izazova i veština, ne samo pripreme i tehničkog izvođenja analize već i primenu kliničkog i medicinskog znanja vezanog za klinički pregled pacijenata, postavljanje indikacija za molekularno genetičke analize i bioinformatičku obradu podataka, interpretaciju i primenu rezultata. Cilj rada: Utvrđivanje genetičke osnove, tačkastih mutacija i malih indel mutacija, analizom kliničkog egzoma metodom sekvenciranja naredne generacije i detekcija mikrodelecija i mikroduplikacija metodom istovremenog umnožavanja vezanih proba kod pacijenata kliničkog genetičara i neuropedijatra sa suspektnim monogenskim oboljenjem i suspektnim mikrodelecijama i mikroduplikacijama. Metod rada: Tokom perioda od 01. avgusta 2021. godine do 01. avgusta 2024. godine u Službi za medicinsku genetiku Instituta za zdravstvenu zaštitu dece i omladine Vojvodine indikovano je od strane kliničkog genetičara 49 analiza kliničkog egzoma metodom sekvenciranja naredne generacije zbog sumnje na monogenske bolesti, a tokom perioda od 01.02.2024. do 01.08.2024. godine, urađeno je 97 analiza metodom istovremenog umnožavanja vezanih proba zbog sumnje na syndrome mikrodelecije i mikroduplikacije, te su u ovom radu navedeni rezultati ispitivanja i dijagnostikovane nasledne bolesti. Rezultati: Analizom kliničkog egzoma metodom sekvenciranja naredne generacije od 49 urađenih analiza detektovani su patološki nalazi – patogene genetičke varijante kod 19 pacijenata tokom perioda od 01. avgusta 2021. godine do 01. avgusta 2024. godine. Analizom istovremenog umnožavanja vezanih proba kod 97 analiziranih pacijenata detektovana je patogena genetička varijanta kod 16 pacijenta, tokom perioda 01.02.2024. do 01.08.2024. godine, te je postignuta visoka stopa detekcije patološkog stanja – obzirom da su ciljano rađena ispitivanja kod pacijenata sa sumnjom na genetički uzrokovanu naslednu bolest, monogensko oboljenje i sindrome mikrodelecija i mikroduplikacija. Zaključak: Uvođenje novih tehnologija u genetički centar Kabineta za molekularnu genetiku i Službu za medicinsku genetiku Instituta za zdravstvenu zaštitu dece i omladine Vojvodine kao što su sekvenciranje naredne generacije i istovremeno umnožavanje vezanih proba su značajni koraci koji vode ka preciznoj medicini i perspektiva su savremene medicine.

 

Ključne reči:

Sekvenciranje naredne generacije, istovremeno umnožavanje vezanih proba, MLPA, retke bolesti, nasledne bolesti, intelektualna ometenost, mikrodelecije, mikroduplikacije, precizna medicina.

 

 

 

Abstract

 

Introduction: The implementation of molecular genetic techniques is a significant event for a genetic centre and represents synchronized processes and a series of complementary activities aimed at introducing new technologies into routine work, as well as overcoming challenges and skills, not only in the preparation and technical performance of the analysis but also in the application of clinical and medical knowledge related to the clinical examination of patients, setting indications for molecular genetic analysis and bioinformatic data processing, interpretation, and application of results. Objective: Determination of the genetic basis, point mutations, and small indel mutations by analysis of the clinical exome using the next-generation sequencing method and detection of microdeletions and icroduplications using the method of multiplex ligation-dependent probe amplification in patients of a clinical geneticist and neuropediatrician with suspected monogenic disease and suspected microdeletions and microduplications. Method: During the period from August 1st, 2021, to August 1st, 2024, in the Medical Genetics Service of the Institute for Health Care of Children and Youth of Vojvodina, a clinical geneticist indicated 49 analyses of the clinical exome using the next-generation sequencing method due to suspicion of monogenic diseases, and during the period from February 1st, 2024, to August 1st, 2024, 97 analyses were performed using the method of multiplex ligation-dependent probe amplification due to suspicion of microdeletion and microduplication syndrome, and this manuscript presented the results of the analyses and diagnosed inherited diseases. Results: Clinical exome analysis using the next-generation sequencing method detected pathological findings in 19 patients out of 49 analyses performed from August 1st, 2021, to August 1st, 2024. Multiplex ligation-dependent probe amplification technique detected in 16 patients a pathogenic genetic variant out of 97 analyzed patients during the period from February 1st, 2024, to August 1st, 2024. A high rate of detection of the pathological condition was achieved, given that the tests were performed specifically in patients with suspected genetic disease, monogenic disease, microdeletion, and microduplication syndromes. Conclusion: The implementation of new technologies in the genetic centre of the Molecular Genetics Cabinet of the Medical Genetics Service of the Institute for Child and Youth Health Protection of Vojvodina, such as next-generation sequencing and multiplex ligation-dependent probe amplification are significant steps leading toward precision medicine and are a perspectivefor modern medicine.

 

Key words:

Next-generation sequencing, Multiplex ligation-dependent probe amplification, MLPA, Rare diseases, Inherited disorders, Intellectual disability, Microdeletions, Microduplications, Precision medicine.

 

 

 

 

References:

1. Hu T, Chitnis N, Monos D, Dinh A. Next-generation sequencing technologies: An overview. Hum Immunol. 2021;82(11):801-811.
2. Yohe S, Thyagarajan B. Review of Clinical Next-Generation Sequencing. Arch Pathol Lab Med. 2017;141(11):1544-1557.
3. Slatko BE, Gardner AF, Ausubel FM. Overview of Next-Generation Sequencing Technologies. Curr Protoc Mol Biol. 2018;122(1):e59.
4. Levy SE, Boone BE. Next-Generation Sequencing Strategies. Cold Spring Harb Perspect Med. 2019;9(7):a025791.
5. Stenzinger A, Moltzen EK, Winkler E, et al. Implementation of precision medicine in healthcare-A European perspective. J Intern Med. 2023;294(4):437-454.
6. Scarano C, Veneruso I, De Simone RR, Di Bonito G, Secondino A, D'Argenio V. The Third-Generation Sequencing Challenge: Novel Insights for the Omic Sciences. Biomolecules. 2024;14(5):568.
7. Köhler S, Gargano M, Matentzoglu N, et al. The Human Phenotype Ontology in 2021. Nucleic Acids Res. 2021;49(D1):D1207-D1217.
8. Bragin E, Chatzimichali EA, Wright CF, Hurles ME, Firth HV, Bevan AP, Swaminathan GJ. DECIPHER: database for the interpretation of phenotype-linked plausibly pathogenic sequence and copy-number variation. Nucleic Acids Res. 2014; 42:D993–D1000.
9. Amberger JS, Bocchini CA, Scott AF. Hamosh A. OMIM.org: leveraging knowledge across phenotype-gene relationships. Nucleic Acids Res. 2019; 47:D1038–D1043.
10. Fu X, Shi Y, Ma J, et al. Advances of multiplex ligation-dependent probe amplification technology in molecular diagnostics. Biotechniques. 2022;73(4):205-213.lpa
11. Petzschner FH. Practical challenges for precision medicine. Science. 2024;383(6679):149-150.
12. Obrenović M, Kavečan I, Privrodski B,  Bojadžijeva Stojanoska B. Molekularno genetska ispitivanja u kliničkoj praksi danas. MD-Medical Data 2017;9(4): 253-256
13. Kavečan I, Nikolić N, Erdeš-Kavečan Đ, Redžek-Mudrinić T, Obrenović M. Značaj precizne medicine i farmakogenomike u preventivnoj pedijatriji. Preventive Paediatrics 2024:10(1-2);060-064.
14. Warburton PE, Sebra RP. Long-Read DNA Sequencing: Recent Advances and Remaining Challenges. Annu Rev Genomics Hum Genet. 2023;24:109-132.
15. Ardui S, Ameur A, Vermeesch JR, Hestand MS. Single molecule real-time (SMRT) sequencing comes of age: applications and utilities for medical diagnostics. Nucleic Acids Res. 2018;46(5):2159-2168.
16. Kumar KR, Cowley MJ, Davis RL. Next-Generation Sequencing and Emerging Technologies. Seminars in thrombosis and hemostasis. 2024;50(7):1026–1038.
    

PDF: 04-Kavečan I. et al MD-Medical Data 2025;17(1) 029-037.pdf