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Moleküler Biyoloji Yöntemlerinin Tıbbi Laboratuvarda Güncel Kullanımı

Year 2025, Volume: 4 Issue: 2, 40 - 53, 30.06.2025

Sinan Tetikoğlu , Funda Bilgili Tetikoğlu , Selcen Çelik Uzuner

https://doi.org/10.59518/farabimedj.1660424

Abstract

Moleküler biyoloji, temelde DNA, RNA ve proteinlerin hücre içi sentezi, işlenmesi ve yıkımına ilişkin hücresel yolaklar ile bu süreçleri düzenleyen mekanizmaların bütününü inceleyen bir bilim dalıdır. Organizmanın en küçük birimi olan hücrelerde bulunan bu moleküllerin tespitine yönelik yöntemlerin klinik uygulamalara aktarılmasıyla, hastalıkların tanı, tedavi ve seyri daha hızlı ve duyarlı yöntemlerle belirlenebilir hale gelmiştir. Günümüzde hemen her hastalığın hücresel bir sebebi ve sonucu olduğu bilinmektedir. Moleküler biyoloji teknikleri, hastalıkların moleküler düzeydeki nedenlerinin aydınlatılması, hastalığa özgü biyobelirteçlerin keşfi ve kişiselleştirilmiş tedavi yaklaşımlarının geliştirilmesi açısından vazgeçilmez araçlardır. Moleküler tekniklerin gelişen teknolojiyle entegrasyonu, bu yöntemleri tanı ve tedavide önemli bir basamak haline getirmiştir. Bu derleme çalışmasının amacı, tıbbi laboratuvarlarda kullanıma uygun moleküler biyoloji yöntemlerine güncel bakış açısı sunmak, son yıllarda yapılan çalışmalardan örnekler vererek bu alandaki ilerlemeleri vurgulamak ve mevcut moleküler biyoloji yöntemlerini tıbbi labortuvarlarda kullanıma entegre etmenin önemine dikkat çekmektir. Ayrıca bu derleme; PCR, NGS, FISH, CRISPR, moleküler blotlama, ELISA ve immünboyama gibi moleküler teknolojilerin tıbbi kullanımlarına, hassasiyetlerine ve klinik uygulamalar üzerindeki katkılarına vurgu yapma yönüyle literatüre katkı sağlayarak, tıbbi laboratuvarlar ve moleküler biyoloji alanlarının ortak paydaşlarına değinmiş olacaktır. Ayrıca moleküler biyoloji tekniklerinin tıbbi laboratuvarlarda kullanılması üzerindeki sınırlılıkları ve gelecek beklentileri de tartışılacaktır.

Keywords

CRISPR FISH Moleküler teknikler NGS PCR Tıbbi laboratuvar

Ethical Statement

Bu makale bir derleme çalışması olduğu için etik kurul onayı gerektirmemektedir.

References

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Current Use of Molecular Biology Methods in Medical Laboratory

Year 2025, Volume: 4 Issue: 2, 40 - 53, 30.06.2025

Sinan Tetikoğlu , Funda Bilgili Tetikoğlu , Selcen Çelik Uzuner

https://doi.org/10.59518/farabimedj.1660424

Abstract

Molecular biology is a scientific discipline that primarily investigates the cellular pathways and regulatory mechanisms involved in the synthesis, processing, and degradation of DNA, RNA, and proteins within the cell. The translation of methods used to detect these molecules in cells—the smallest functional units of the organism—into clinical applications has enabled the diagnosis, treatment, and monitoring of diseases through more rapid and sensitive techniques. Today, it is well established that nearly every disease has a cellular cause and consequence. Molecular biology techniques are indispensable tools for elucidating the molecular basis of diseases, discovering disease-specific biomarkers, and developing personalized therapeutic strategies. The integration of molecular techniques with advancing technology has made these methods a critical step in diagnosis and therapy. This review aims to provide an up-to-date perspective on molecular biology techniques suitable for use in medical laboratories, highlight recent advances in the field by presenting examples from recent studies, and emphasize the importance of integrating current molecular biology methods into medical laboratory practice. Additionally, this review seeks to contribute to the literature by discussing the medical applications, sensitivities, and clinical impacts of key molecular technologies such as PCR, NGS, FISH, CRISPR, molecular blotting, ELISA, and immunostaining. Furthermore, the limitations of employing molecular biology techniques in medical laboratories and future expectations for the field will also be addressed, offering a comprehensive evaluation for stakeholders across both molecular biology and medical laboratory sciences.

Keywords

CRISPR FISH Medical laboratory Molecular techniques NGS PCR

References

  • Speers DJ. Clinical applications of molecular biology for infectious diseases. Clin Biochem Rev. 2006;27(1):39-51.
  • Arikat S, Saboor M. Evolving role of clinical laboratories in precision medicine: a narrative review. J Lab Precis Med 2024;9:17.
  • Delidow BC, Lynch JP, Peluso JJ, White BA. Polymerase chain reaction : basic protocols. Methods Mol Biol. 1993;15:1-29.
  • Khehra N, Padda IS, Swift CJ. Polymerase Chain Reaction (PCR). Updated March 6, 2023. In: StatPearls [Internet]. Treasure Island (FL): StatPearls Publishing; January 2025–. Accessed March 6, 2025. https://www.ncbi.nlm.nih.gov/books/NBK589663
  • Zhu H, Zhang H, Xu Y, Laššáková S, Korabečná M, Neužil P. PCR past, present and future. Biotechniques. 2020;69(4):317-325.
  • Artika IM, Dewi YP, Nainggolan IM, Siregar JE, Antonjaya U. Real-Time Polymerase Chain Reaction: Current techniques, applications, and role in COVID-19 Diagnosis. Genes (Basel). 2022;13(12):2387.
  • Navarro E, Serrano-Heras G, Castaño MJ, Solera J. Real-time PCR detection chemistry. Clin Chim Acta. 2015;439:231-250.
  • Dutta D, Naiyer S, Mansuri S, et al. COVID-19 diagnosis: A comprehensive review of the RT-qPCR Method for Detection of SARS-CoV-2. Diagnostics (Basel). 2022;12(6):1503.
  • Sugita S, Takase H, Nakano S. Practical use of multiplex and broad-range PCR in ophthalmology. Jpn J Ophthalmol. 2021;65(2):155-168.
  • Nyaruaba R, Mwaliko C, Dobnik D, et al. Digital PCR applications in the SARS-CoV-2/COVID-19 Era: a roadmap for future outbreaks. Clin Microbiol Rev. 2022;35(3):e00168-21.
  • Yugovich O, Bunce M, Harbison SA. Point-of-need species identification using non-PCR DNA-based approaches to combat wildlife crime. Forensic Sci Int Genet. 2025;78:103278.
  • Lee CL, Chuang CK, Chiu HC, et al. Understanding Genetic Screening: Harnessing Health Information to Prevent Disease Risks. Int J Med Sci. 2025;22(4):903-919.
  • Grody WW, Dunkel-Schetter C, Tatsugawa ZH, et al. PCR-based screening for cystic fibrosis carrier mutations in an ethnically diverse pregnant population. Am J Hum Genet. 1997;60(4):935-947.
  • Stern HJ. Preimplantation Genetic Diagnosis: Prenatal Testing for Embryos Finally Achieving Its Potential. J Clin Med. 2014;3(1):280.
  • Satam H, Joshi K, Mangrolia U, et al. Next-Generation Sequencing Technology: Current Trends and Advancements. Biology (Basel). 2023;12(7):997.
  • Katara A, Chand S, Chaudhary H, Chaudhry V, Chandra H, Dubey RC. Evolution and applications of next generation sequencing and its intricate relations with chromatographic and spectrometric techniques in modern day sciences. Journal of Chromatography Open. 2024;5:100121.
  • Ghoreyshi N, Heidari R, Farhadi A, et al. Next-generation sequencing in cancer diagnosis and treatment: clinical applications and future directions. Discov Oncol. 2025;16(1):578.
  • Salk JJ, Schmitt MW, Loeb LA. Enhancing the accuracy of next-generation sequencing for detecting rare and subclonal mutations. Nat Rev Genet. 2018;19(5):269.
  • Kamps R, Brandão RD, van den Bosch BJ, et al. Next-Generation Sequencing in Oncology: Genetic Diagnosis, Risk Prediction and Cancer Classification. Int J Mol Sci. 2017;18(2):308.
  • Wang Y, Yang Q, Wang Z. The evolution of nanopore sequencing. Front Genet. 2015;5:449.
  • Wang Y, Zhao Y, Bollas A, Wang Y, Au KF. Nanopore sequencing technology, bioinformatics and applications. Nat Biotechnol. 2021;39(11):1348-1365.
  • Shakoori AR. Fluorescence In Situ Hybridization (FISH) and Its Applications. Chromosome Structure and Aberrations. 2017;343-367.
  • Jensen E. Technical review: In situ hybridization. Anat Rec (Hoboken). 2014;297(8):1349-1353.
  • Harun A, Liu H, Song S, et al. Oligonucleotide Fluorescence In Situ Hybridization: An Efficient Chromosome Painting Method in Plants. Plants (Basel). 2023;12(15):2816.
  • Kudman S, Semaan A, Assaad MA, et al. Optimization of Fluorescence In Situ Hybridization Protocols in the Era of Precision Medicine. Curr Protoc. 2024;4(6):e1093.
  • Cui C, Shu W, Li P. Fluorescence In situ Hybridization: Cell-Based Genetic Diagnostic and Research Applications. Front Cell Dev Biol. 2016;4:89.
  • Mohamed AM, Eid M, Eid O, et al. Generation of Dual-Color FISH probes targeting 9p21, Xp21, and 17p13.1 loci as diagnostic markers for some genetic disorders and cancer in Egypt. J Genet Eng Biotechnol. 2025;23(1):100449.
  • Yang T, Kang L, Li D, Song Y. Immunotherapy for HER-2 positive breast cancer. Front Oncol. 2023;13:1097983.
  • Tommasi C, Airò G, Pratticò F, et al. Hormone Receptor-Positive/HER2-Positive Breast Cancer: Hormone Therapy and Anti-HER2 Treatment: An Update on Treatment Strategies. J Clin Med. 2024;13(7):1873.
  • Baez-Navarro X, Groenendijk FH, Oudijk L, et al. HER2-low across solid tumours: different incidences and definitions. Pathology. 2025;57(4):403-414.
  • Rose NC, Barrie ES, Malinowski J, et al. Systematic evidence-based review: The application of noninvasive prenatal screening using cell-free DNA in general-risk pregnancies [published correction appears in Genet Med. 2022 Sep;24(9):1992.
  • Xu Y, Li Z. CRISPR-Cas systems: Overview, innovations and applications in human disease research and gene therapy. Comput Struct Biotechnol J. 2020;18:2401-2415.
  • Lone BA, Karna SKL, Ahmad F, Shahi N, Pokharel YR. CRISPR/Cas9 System: A bacterial tailor for genomic engineering. Genet Res Int. 2018;2018:3797214.
  • Redman M, King A, Watson C, King D. What is CRISPR/Cas9? Arch Dis Child Educ Pract Ed. 2016;101(4):213.
  • Jiang Y, Qian F, Yang J, et al. CRISPR-Cpf1 assisted genome editing of Corynebacterium glutamicum. Nature Communications 2017 8:1. 2017;8(1):1-11.
  • Li T, Yang Y, Qi H, et al. CRISPR/Cas9 therapeutics: progress and prospects. Signal Transduct Target Ther. 2023;8(1):36.
  • Allemailem KS, Almatroodi SA, Almatroudi A, et al. Recent advances in genome-editing technology with CRISPR/Cas9 variants and stimuli-responsive targeting approaches within tumor cells: A future perspective of cancer management. Int J Mol Sci. 2023;24(8):7052.
  • Khoshandam M, Soltaninejad H, Mousazadeh M, Hamidieh AA, Hosseinkhani S. Clinical applications of the CRISPR/Cas9 genome-editing system: Delivery options and challenges in precision medicine. Genes Dis. 2024;11(1):268-282.
  • Mustafa MI, Makhawi AM. SHERLOCK and DETECTR: CRISPR-Cas systems as potential rapid diagnostic tools for emerging infectious diseases. J Clin Microbiol. 2021;59(3):e00745-20.
  • Ebrahimi S, Khanbabaei H, Abbasi S, et al. CRISPR-Cas System: A Promising Diagnostic Tool for Covid-19. Avicenna J Med Biotechnol. 2022;14(1):3.
  • Tanaka PP, Monteiro CJ, Duarte MJ, et al. The CRISPR-Cas9 system is used to edit the autoimmune regulator gene in vitro and in vivo. Adv Exp Med Biol. 2025;1471:269-283.
  • Li X, Wang Z, Man X, Dai X, Zhou Q, Zhang S. Research advances CRISPR gene editing technology generated models in the study of epithelial ovarian carcinoma. Gynecol Oncol. 2025;195:34-44.
  • Fang S, Wang G, Yang LH. [Advances in AAV-CRISPR/Cas9-Mediated Hemophilia A Gene Therapy --Review]. Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2023;31(6):1890-1893.
  • Lin YT, Seo J, Gao F, et al. APOE4 causes widespread molecular and cellular alterations associated with alzheimer’s disease phenotypes in human iPSC-derived brain cell types. Neuron. 2018;98(6):1141-1154.e7.
  • Thapar N, Eid MAF, Raj N, Kantas T, Billing HS, Sadhu D. Application of CRISPR/Cas9 in the management of Alzheimer’s disease and Parkinson’s disease: a review. Ann Med Surg (Lond). 2023;86(1):329-335.
  • Ciafaloni E, Kumar A, Liu K, et al. Age at onset of first signs or symptoms predicts age at loss of ambulation in Duchenne and Becker Muscular Dystrophy: Data from the MD STARnet. J Pediatr Rehabil Med. 2016;9(1):5-11.
  • Ou L, Przybilla MJ, Tăbăran AF, et al. A novel gene editing system to treat both Tay-Sachs and Sandhoff diseases. Gene Ther. 2020;27(5):226-236.
  • Kang Y, Li H, Liu Y, Li Z. Regulation of VEGF-A expression and VEGF-A-targeted therapy in malignant tumors. J Cancer Res Clin Oncol. 2024;150(5):221.
  • Dourthe ME, Baruchel A. CAR T-cells for acute leukemias in children: current status, challenges, and future directions. Cancer Metastasis Rev. 2025;44(2):47.
  • Tariq H, Khurshid F, Khan MH, et al. CRISPR/Cas9 in the treatment of sickle cell disease (SCD) and its comparison with traditional treatment approaches: a review. Ann Med Surg (Lond). 2024;86(10):5938-5946.
  • Weaver SB, Singh D, Wilson KM. Gene Therapies for Sickle Cell Disease. J Pharm Technol. 2024;40(5):236-247.
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There are 75 citations in total.

Details

Primary Language Turkish
Subjects Medical Biochemistry and Metabolomics (Other), Health Services and Systems (Other)
Journal Section Reviews
Authors

Sinan Tetikoğlu 0000-0002-7725-6034

Funda Bilgili Tetikoğlu 0000-0003-3734-4803

Selcen Çelik Uzuner 0000-0002-9558-7048

Early Pub Date June 29, 2025
Publication Date June 30, 2025
Submission Date March 19, 2025
Acceptance Date May 14, 2025
Published in Issue Year 2025 Volume: 4 Issue: 2

Cite

AMA Tetikoğlu S, Bilgili Tetikoğlu F, Çelik Uzuner S. Moleküler Biyoloji Yöntemlerinin Tıbbi Laboratuvarda Güncel Kullanımı. Farabi Med J. June 2025;4(2):40-53. doi:10.59518/farabimedj.1660424
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