Bioscience Methods 2024, Vol.15, No.3, 91-101 http://bioscipublisher.com/index.php/bm 92 molecular diagnostics in detecting infectious diseases in pets has led to better management and control of these diseases, ultimately improving animal health and welfare (Balamurugan et al., 2010; Granberg et al., 2014). This study explores the advancements in molecular diagnostics and their impact on pet disease detection; provides a comprehensive overview of the current state of molecular diagnostics in veterinary medicine, and highlights the benefits and challenges associated with these techniques. By examining various molecular diagnostic methods and their applications, this study seeks to underscore the importance of early and accurate diagnosis in improving the health outcomes of pets. The scope includes an analysis of different molecular diagnostic tools, their clinical applications, and the future prospects of molecular diagnostics in veterinary medicine. 2 Fundamentals of Molecular Diagnostics 2.1 Definition and key concepts Molecular diagnostics refers to a collection of techniques used to analyze biological markers in the genome and proteome. These techniques are employed to diagnose and monitor diseases, detect risk, and decide which therapies will work best for individual patients. The key concepts include the detection and quantification of nucleic acids (DNA and RNA), proteins, and other molecules that provide information about the health status of an organism. 2.2 Types of molecular diagnostic techniques Polymerase Chain Reaction (PCR) is a widely used method in molecular diagnostics due to its high sensitivity and specificity. PCR amplifies small segments of DNA or RNA, making it possible to detect even minute quantities of pathogens or genetic mutations. This technique has been extensively applied in various fields, including dentistry, where it helps in the detection and identification of microorganisms responsible for periodontal and endodontic infections (Shahi et al., 2018). PCR's ability to provide rapid and accurate results makes it a cornerstone in the diagnosis of infectious diseases (Kurkela and Brown, 2009). Next-Generation Sequencing (NGS) is a powerful tool that allows for the comprehensive analysis of genetic material. NGS can sequence entire genomes or specific areas of interest, providing detailed information about genetic variations and mutations. This technology has revolutionized the field of molecular diagnostics by enabling the identification of pathogens and genetic disorders with high precision. NGS is particularly useful in the investigation of fevers of unknown origin (FUO) and has the potential to replace traditional microbial identification methods (Wright et al., 2021). Microarrays are another important molecular diagnostic technique that allows for the simultaneous analysis of thousands of genetic sequences. This method is highly sensitive and can detect multiple pathogens in a single test. However, the high cost and complexity of microarrays limit their routine use in clinical diagnostics. Despite these limitations, microarrays hold promise for the future of molecular diagnostics, especially in the detection of genetic polymorphisms and immune markers (Kurkela and Brown, 2009). CRISPR-based diagnostics represent a cutting-edge approach in molecular diagnostics. Leveraging the CRISPR-Cas system, these techniques offer high sensitivity and specificity for nucleic acid detection. CRISPR diagnostics are particularly advantageous for point-of-care testing due to their simplicity and rapid turnaround time. They have been successfully used to detect various pathogens, including SARS-CoV-2, and have shown potential for use in low-resource settings (Figure 1) (Hatoum-Aslan, 2018; Kaminski et al., 2021; Abudayyeh and Gootenberg, 2021). The development of CRISPR-based tools continues to expand, providing new opportunities for accurate and accessible disease detection (Yin et al., 2020; Weng et al., 2023). 2.3 Advantages over traditional methods Molecular diagnostic techniques offer several advantages over traditional methods. They provide higher sensitivity and specificity, allowing for the detection of low levels of pathogens or genetic mutations. These techniques also offer faster turnaround times, which is crucial for timely diagnosis and treatment. Additionally, molecular diagnostics can be performed on a variety of sample types, including blood, tissue, and swabs, making them versatile tools in disease detection. The ability to perform multiplex testing, where multiple
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