A deadly and chaotic virus named SARS-CoV-2 has become a big threat causing acute respiratory distress, which has brought an unprecedented challenge to our healthcare system. This, in turn, arouses the need for on-site, sensitive, specific, and rapid diagnostic tests that can accurately detect pathogens. This was the time when care providers worked in an integrated manner to develop a nucleic acid testing kit.
Figure 1. Children’s Case of Respiratory Infection
How Does Nucleic Acid Diagnostic Kit Detect and Identify Six Respiratory Pathogens?
The nucleic acid diagnostic kit detects and identifies six common bacteria: Klebsiella pneumonia, Haemophilus influenza, Staphylococcus aureus, Streptococcus pneumonia, Pseudomonas aeruginosa, Legionella pneumophila that can cause lower respiratory tract infections.
- Protocol to detect and identify these bacteria
To detect and identify these bacteria, DNA or RNA extraction and purification from bacteria were performed using a nucleic acid extraction kit.
Figure 2. Sample collection procedure for the DNA test
2. DNA or RNA extraction
For the extraction of RNA or DNA, the spiked sample is added to a tube to extract RNA or DNA for subsequent analyses.
3. Nucleic acid amplification
There are several ways of nucleic acid amplification: polymerase chain reaction (PCR) amplification, transcription-mediated amplification (TMA), or strand displacement amplification. Compared with other nucleic acid amplification methods, the multiplex-based PCR kits bring accuracy, convenience, and higher throughput to the test.
Figure 3. Sansure Six Respiratory Pathogens Nucleic Acid Diagnostic Kit (PCR-Fluorescence Probing)
Multiplex-Based Nucleic Acid Diagnosis
As aforementioned, the diagnosis of respiratory infection can be performed using multiplex-based PCR kits, which detect multiple targets in one tube simultaneously. Sansure innovates multiplex-based PCR kits by integrating fluorescence color and melting temperature as a two-dimensional label for homogeneous multiplex PCR detection, which is highly accurate and efficient.
For example, in two different clinical studies, to evaluate the clinical performance of Sansure six respiratory pathogens diagnostic kit, a total of 2047 samples were tested in which 1548 samples were tested in the first trial. From the results it was observed that overall sample consistency percentage was 99.88% (99.31% -100.00%), and negative consistency percentage was 97.72% (95%CI: 96.37% – 98.66%), and 98.84% (95%CI: 98.17% – 99.31%). The sensitivity ranged from 100% [95%CI: 99.32%-100.0%] for bacteria and overall specificity was excellent (72.22% [95%CI: 69.35%-74.97%]).
Advantages of Multiplex PCR Diagnostic Kit for Lower Respiratory Infection
Multiplex PCR technology is increasingly used to diagnose respiratory bacteria partly because of its superior sensitivity, relatively rapid turnaround time, and ability to detect multiple pathogens in a single test. Taking Sansure nucleic acid diagnostic kits as an example, the benefits brought by multiplex PCR diagnostic kits for lower respiratory infection can be:
Respiratory illnesses share similar symptoms, making correct diagnosis complicated. However, multiplex PCR is able to match the performance of quantitative bacterial culture and provide greater specificity, thus increasing the accuracy of data analysis.
The time for the expense of reagents and preparation is reduced in multiplex PCR than in other methods where several tubes of uniplex PCRs are used. Multiplex PCR enables the operator to gain more information with the limited starting material. It is time-saving, cost-effective, and brings higher throughput, making it ideal for repeated analysis of similar types of samples.
3. Medication Guidance
Multiplex PCR allows the correct use of antimicrobial drugs, including antibiotics and anti-influenza agents (as detailed earlier), as it can be used simultaneously with platforms that allow the identification of antimicrobial resistance to commonly used drugs.
It is highly likely that multiplex PCR platforms, particularly those with high sensitivity and specificity and with low turnaround time, will become essential when new drugs that are effective and safe against most respiratory viruses are available.
Figure 4. Medication for Lower Respiratory Infection
Sansure nucleic acid diagnostic kit uses advanced magnetic beads technology to carry out room temperature lysis with significantly less contamination with a capacity of up to 96 samples at one time with a sensitivity of 2.0 TCID50/mL for Influenza A and Influenza B virus. 500.0 copies/mL for Respiratory syncytial virus, Adenovirus, Mycoplasma pneumonia and Human rhinovirus to enhance large-scale screening efficiency.
The enterprise reference for the limit of detection of this kit is 900 CFU/mL (klebsiella pneumoniae), 15 CFU/mL (streptococcus pneumoniae), 625 CFU/mL (haemophilus influenzae), 675 CFU/mL (pseudomonas aeruginosa), 340 CFU/mL (legionella pneumophila), 2875 CFU/mL (staphylococcus aureus).
In addition, it can effectively aid clinicians in making an accurate diagnosis, detecting the pathogens that cause symptoms, and formulating treatment plans.
For more information, don’t hesitate to get in touch with Sansure at: https://www.sansureglobal.com/.