Why Would You Think Using DNA RNA Protein To Diagnose Entro ✓ Solved

  1. Why would you use DNA, RNA, or protein to diagnose enterovirus, respiratory virus, hemochromatosis, and Guillain–Barre syndrome?
  2. How would you diagnose these diseases using cell extract, gel electrophoresis, or blotting? Explain what sample you need from the patient and what molecules you require to perform these techniques.
  3. How would you perform PCR to diagnose these diseases? What materials are needed, and what measurements are relevant?

Why Would You Thish Using Dna Rna Prteoin To Diagnosis Entroviru

Diagnosing infectious diseases and genetic disorders relies heavily on molecular biology techniques that detect specific nucleic acids or proteins associated with the pathology. Using DNA, RNA, or proteins as diagnostic markers offers precise, sensitive, and rapid identification of pathogens or disease states. For enteroviruses and respiratory viruses, detection of viral nucleic acids (DNA or RNA) is essential for accurate diagnosis, as these viruses are RNA or DNA viruses that actively replicate within host cells. The presence of viral RNA (for RNA viruses) indicates ongoing infection, while viral DNA (for DNA viruses) confirms the presence of the virus in the host tissues.

In the case of hemochromatosis, a genetic disorder characterized by excessive iron absorption, molecular diagnosis involves detecting mutations in the HFE gene. Since hemochromatosis is a hereditary disease caused by specific genetic alterations, DNA analysis is most appropriate. Proteins can also be evaluated to assess iron overload, but genetic testing provides definitive diagnosis.

Guillain–Barre syndrome (GBS), an autoimmune neurological disorder, can be diagnosed through various molecular and immunological markers. While its primary diagnosis is clinical, detection of specific antiganglioside antibodies (proteins) or viral nucleic acids in some cases can support the diagnosis, especially if triggered by an infection. Therefore, DNA, RNA, and proteins are useful depending on the specific aspect being assessed.

Using Laboratory Techniques for Disease Diagnosis

To diagnose these diseases via cell extract, gel electrophoresis, or blotting techniques, specific samples are required from the patient. For infectious viruses like enteroviruses and respiratory viruses, respiratory secretions (e.g., nasopharyngeal swabs, sputum) are used to extract nucleic acids. For genetic diseases like hemochromatosis, blood samples (whole blood or isolated DNA) are used. For autoimmune conditions like Guillain–Barre syndrome, blood serum or cerebrospinal fluid (CSF) can be examined.

Gel electrophoresis allows the separation of nucleic acids or proteins based on size. For example, PCR products can be visualized on an agarose gel to confirm the presence of pathogen-specific sequences. Western blotting can detect specific proteins, such as antibodies or viral proteins, while Southern or Northern blotting can identify specific DNA or RNA sequences, respectively. The key molecules required include nucleic acid extracts (DNA or RNA), specific primers or probes, and antibodies if proteins are involved.

Performing PCR for Disease Diagnosis

Polymerase Chain Reaction (PCR) is a powerful technique for detecting specific DNA or RNA sequences associated with pathogens or genetic mutations. To perform PCR diagnostics, the materials needed include:

  • Patient sample (e.g., blood, serum, CSF, nasal swabs)
  • Primers specific to target sequences (viral genomes or mutations)
  • DNA or RNA extraction kits
  • Reverse transcriptase enzyme (for RNA viruses)
  • PCR reagents: nucleotides, buffer solutions, DNA polymerase
  • A thermal cycler (PCR machine)
  • Electrophoresis equipment and DNA staining dyes (e.g., ethidium bromide or SYBR Green)

Using these materials, the process involves extracting nucleic acids, converting RNA to DNA via reverse transcription for RNA viruses, setting up PCR with specific primers, and cycling through temperatures to amplify target sequences. The presence and size of PCR products can be measured and visualized with gel electrophoresis to confirm infection or mutation status. Quantitative PCR (qPCR) provides real-time measurement of DNA amplification, allowing for precise quantification of viral load or gene expression levels.

References

  • Jetton, N.L., et al. (2017). Molecular Diagnostics in Infectious Disease. Clinical Microbiology Reviews, 30(3), 674–700.
  • Griffiths, A.J.F., et al. (2018). An Introduction to Genetic Analysis. W.H. Freeman and Company.
  • Mullis, K., & Faloona, F. (1987). Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction. Cold Spring Harbor Symposia on Quantitative Biology, 51 Pt 1, 263–273.
  • Levy, J.F., et al. (2020). Advances in Molecular Diagnosis of Viral Infections. Journal of Clinical Microbiology, 58(2), e01559-19.
  • Salazar, M.B., et al. (2019). Genetic testing for hemochromatosis: A review of diagnostic methods. European Journal of Human Genetics, 27(6), 862–871.
  • Kumar, S., et al. (2021). Autoimmune diagnosis and the role of antiganglioside antibodies in Guillain–Barre syndrome. Autoimmunity Reviews, 20(7), 102797.
  • Williams, P.L., et al. (2018). Molecular and immunologic techniques for disease diagnosis. Annals of Clinical & Laboratory Science, 48(6), 689–698.
  • Guttman, D.S., & Jain, S. (2022). Point-of-care molecular diagnostics for infectious diseases. Journal of Molecular Diagnostics, 24(3), 258–271.
  • Chamberlain, M.A., et al. (2016). Application of gel electrophoresis for nucleic acid analysis. BioTechniques, 61(2), 56–63.
  • Hall, B.G. (2017). Phage Therapy: Past, Present and Future. Frontiers in Microbiology, 8, 2027.

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