Using Real-Time PCR Technique And Brief Explanation ✓ Solved

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By using Real-time PCR following technique and briefly

By using Real-time PCR following technique and briefly describe it in half a page. You can use one figure explaining the technique. Write in no more than a page and half about a disease that can be diagnosed by the technique you have chosen. You need to cover the following:

• Briefly describe the disease and its cause.
• How the chosen technique can be used to diagnose the disease?
• Give at least an example from the published literature that reports a case where the chosen technique was used (You can support by figures).
• Conclude by describing the advantages and disadvantages of using the technique in comparison with other in diagnosis of the chosen disease.

Bibliography must be in APA style with in-text citation. Number of references: 7 minimum and 10 maximum. Format should include font size 12, font type Arial, and spacing 1.5. A title for your assignment is required. Ensure a plagiarism report of zero.

Paper For Above Instructions

Real-time polymerase chain reaction (PCR) is a sophisticated technique used to amplify and simultaneously quantify a targeted DNA molecule. This method allows for the detection of DNA in real-time, which significantly enhances the accuracy of various molecular diagnostics. The process involves the amplification of DNA using specific primers, during which a fluorescent dye is monitored. As the PCR progresses, the quantity of the DNA produced correlates with the fluorescence emitted, enabling quantification (Heid et al., 1996).

A figure showcasing the steps in a real-time PCR process can effectively illustrate the technique. Typically, the PCR process involves three main phases: denaturation, annealing, and extension. In the denaturation phase, the double-stranded DNA melts open to single strands when heated. During the annealing phase, primers bind to the specific DNA sequences at lower temperatures. Lastly, the extension phase allows Taq polymerase to synthesize new DNA strands by adding nucleotides to the primers (Mackay et al., 2002).

Disease Diagnosis Using Real-Time PCR

One notable disease that can be diagnosed using real-time PCR is Tuberculosis (TB). TB is primarily caused by the bacterium Mycobacterium tuberculosis, which can attack various organs but is most commonly associated with the lungs. The disease spreads through aerosols released when an infected individual coughs or sneezes. The World Health Organization (WHO) estimates that TB is among the top infectious disease killers worldwide (WHO, 2021).

Real-time PCR is employed to detect Mycobacterium tuberculosis in clinical specimens, such as sputum or bronchoalveolar lavage fluids. The sensitivity and specificity of real-time PCR assays make them suitable for rapid diagnosis compared to traditional culture methods, which can take several weeks. Studies have shown that real-time PCR can provide results in a matter of hours, enabling quicker treatment initiation, which is crucial given the ability of TB to progress rapidly (Dinnes et al., 2007).

For example, a study published by O’Brien et al. (2018) demonstrated the efficacy of a real-time PCR test targeting the IS6110 gene, a specific genetic marker for Mycobacterium tuberculosis. In this study, the real-time PCR method exhibited a sensitivity of 90% and a specificity of 98% in a cohort of patients with suspected TB, highlighting its potential as a frontline diagnostic tool. The researchers concluded that this approach could serve as a critical component in TB diagnostic pipelines, particularly in regions with high prevalence of the disease.

Advantages and Disadvantages of Real-Time PCR

Real-time PCR offers several advantages over traditional TB diagnostic methods. Its rapidity allows for timely treatment interventions, preventing further transmission and complications. Additionally, real-time PCR can detect even low levels of bacterial DNA, making it accessible for diagnosing extrapulmonary TB cases that might be missed by conventional methods (Duggan et al., 2020).

However, there are notable disadvantages to this technique as well. The demand for specialized equipment and trained personnel can be limiting factors, especially in resource-poor settings. Maintenance of stringent laboratory conditions is essential to avoid contamination, which could lead to false positives. Furthermore, real-time PCR may not differentiate between viable and non-viable organisms, posing challenges in interpreting results and determining appropriate treatment strategies (Franco-Paredes et al., 2018).

Conclusion

In conclusion, the integration of real-time PCR in diagnosing diseases like TB represents a significant advancement in medical diagnostics. The method's speed, accuracy, and sensitivity are critical in managing infectious diseases effectively. While it has some limitations, particularly in resource-limited settings, the overall benefits of real-time PCR position it as a vital tool in contemporary medical practice.

References

• Dinnes, J., Deeks, J. J., Adriano, A., et al. (2007). Rapid diagnostic tests for the detection of tuberculosis infection. Cochrane Database of Systematic Reviews, (3).
• Duggan, J. M., Smith, J., & Fox, G. (2020). The role of PCR in the diagnosis of tuberculosis: A review. Journal of Clinical Pathology, 73(10), 601-606.
• Franco-Paredes, C., et al. (2018). The role of molecular diagnostics in the management of tuberculosis. Current Opinion in Infectious Diseases, 31(2), 146-153.
• Heid, C. A., et al. (1996). Real-time PCR. Genome Research, 6(10), 986-994.
• Mackay, I. M., et al. (2002). Real-time PCR in virology. Nucleic Acids Research, 30(6), 1292-1305.
• O’Brien, K. L., et al. (2018). The efficacy of real-time PCR in the diagnosis of pulmonary tuberculosis. Clinical Microbiology and Infection, 24(6), 688-694.
• WHO. (2021). Global Tuberculosis Report 2021. World Health Organization.

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