What is Sepsis? How to Use Sepsis Real-Time qPCR Panel Detection Kit?

Sepsis is a medical emergency that claims millions of lives worldwide each year, yet it remains relatively unknown to the general public. Often referred to as “blood poisoning,” sepsis is a condition that can affect anyone, from infants to the elderly, and can arise from seemingly minor infections or injuries. In this blog, we will delve into what sepsis is, its causes, symptoms, and the importance of early detection and treatment.

What is Sepsis?

Sepsis is a severe and potentially life-threatening condition that occurs when the body’s response to infection goes fail. Normally, when you get an infection, your body’s immune system fights off the invading pathogens like bacteria, viruses, or fungi [1]. However, in sepsis, the body’s immune response becomes overly aggressive, triggering widespread inflammation that can harm healthy tissues and organs [1].

   

Sepsis can quickly progress to severe sepsis or septic shock, where the body’s organs, such as the heart, lungs, kidneys, liver, and brain, can start to fail. If left untreated, the disease can lead to multiple organ failure and death.

What are the Causes of Sepsis?

Sepsis can be caused by a variety of infections, including:

1.Bacterial Infections: Most cases of sepsis are caused by bacterial infections. Common sources include urinary tract infections, pneumonia, and skin infections [2].

2.Viral Infections: While less common, viral infections like COVID-19 can also lead to sepsis [2].

3.Fungal Infections: Fungal sepsis is rarer but can occur in individuals with weakened immune systems [2].

4.Infections from Medical Procedures: In some cases, sepsis can develop after surgical procedures or the insertion of medical devices like catheters [2].

What are the Symptoms of Sepsis?

Sepsis can be challenging to diagnose, as its symptoms can mimic those of other illnesses. Some common signs of sepsis include:

• Fever or Hypothermia: A high fever or abnormally low body temperature.
• Rapid Heart Rate: An elevated heart rate, often exceeding 90 beats per minute.
• Rapid Breathing: An increased respiratory rate, typically over 20 breaths per minute.
• Confusion or Altered Mental State: Difficulty thinking clearly or changes in consciousness.
• Low Blood Pressure: A drop in blood pressure that may lead to dizziness or fainting.
• Decreased Urine Output: A significant decrease in the amount of urine produced.
• Skin Changes: Skin may appear mottled or discolored.

It’s crucial to recognize that sepsis can develop rapidly, and early detection is key to improving the chances of survival.

Risk Groups

Sepsis can affect anyone, regardless of their level of health or physical fitness, as well as their geographical location. However, some individuals face an elevated risk, including:

• Infants under the age of 1.
• Individuals aged 60 and older.
• People who have had their spleen removed.
• Those with chronic conditions such as lung, liver, or heart diseases.
• Individuals with compromised immune systems, such as those with AIDS or diabetes.

What are the Diagnosis Methods and Treatment for Sepsis?

Diagnosing sepsis can be challenging because its symptoms can mimic those of other medical conditions. Healthcare providers typically use a combination of clinical assessments and laboratory tests to diagnose sepsis. Here are some common methods and approaches used in the diagnosis of sepsis:

1.Clinical Evaluation: Healthcare professionals assess the patient’s medical history and perform a physical examination. They look for signs such as fever or hypothermia, rapid heart rate, rapid breathing, altered mental state, and low blood pressure [3]. The presence of these clinical signs may raise suspicion of sepsis.

2.Blood Tests: Blood tests are crucial for diagnosing sepsis. They can include:

• Complete Blood Count (CBC): This test measures the levels of various blood components, including white blood cells [3]. A significant increase or decrease in white blood cell count can indicate an infection.
• Blood Cultures: These tests involve taking samples of blood to identify the specific bacteria or other pathogens causing the infection.
• Lactate Levels: Elevated lactate levels in the blood may suggest tissue damage or oxygen deprivation, both of which can occur in sepsis [3].

3.Inflammatory Marker Tests: Measurement of inflammatory markers like C-reactive protein (CRP) and procalcitonin can help assess the body’s response to infection and inflammation [3]. Elevated levels may indicate sepsis.

 

4.Polymerase Chain Reaction (PCR): PCR can be used to detect the presence of specific pathogens, such as bacteria or viruses, by amplifying and identifying their genetic material (DNA or RNA). PCR is particularly useful for identifying the causative agents of infections associated with sepsis.

5.Imaging Studies: In some cases, imaging tests such as X-rays, CT scans, or ultrasounds may be used to identify the source of infection or evaluate the condition of affected organs.

6.Microbiological Cultures: Cultures of samples from suspected infection sites (e.g., urine, sputum, wound, or cerebrospinal fluid) may be performed to identify the specific microorganism causing the infection [3].

7.Arterial Blood Gas Analysis: This test can help assess the patient’s oxygen levels and acid-base balance, which can be disrupted in severe cases of sepsis [3].

8.Septic Screen: A combination of tests may be performed to evaluate markers of sepsis, including white blood cell count, lactate, and procalcitonin levels.

It’s essential to diagnose sepsis as early as possible to initiate prompt treatment, as delays can significantly impact the patient’s prognosis. The combination of clinical assessment, blood tests, and other diagnostic methods helps healthcare providers make an accurate diagnosis and tailor treatment accordingly. Early recognition and intervention are critical for improving outcomes in sepsis cases.

Treatment typically involves antibiotics to combat the underlying infection, intravenous fluids to maintain blood pressure, and other supportive measures to stabilize vital functions. In severe cases, patients may require admission to an intensive care unit (ICU) and additional therapies, such as mechanical ventilation or kidney dialysis.

Preventing Methods

While sepsis can strike anyone, there are steps you can take to reduce your risk:

• Practice Good Hygiene.
• Stay Up-to-Date on Vaccinations
• Manage Chronic Illnesses
• Promptly Treat Infections
• Vaccination

How to Use Sepsis Real-Time qPCR Panel Detection Kit?

Using a Sepsis Real-Time qPCR Panel Detection Kit involves several steps to accurately detect and identify pathogens associated with sepsis using quantitative polymerase chain reaction (qPCR). Please note that specific kit instructions may vary slightly depending on the manufacturer, so always refer to the kit’s user manual provided with the product for precise guidelines. Here’s a general overview of how to use such a kit:

1. Sample Collection and Preparation: Collect clinical samples (e.g., blood, urine, cerebrospinal fluid) from the patient as per standard medical procedures. Follow recommended guidelines for sample collection and handling to prevent contamination.
2. Nucleic Acid Extraction (if required): Some kits may require you to extract nucleic acids (DNA or RNA) from the collected samples before PCR. Follow the manufacturer’s instructions for nucleic acid extraction if this step is necessary.
3. Setting Up the qPCR Reaction: Prepare the qPCR reaction mix according to the kit’s instructions. Typically, this involves combining the kit components (primers, probes, enzymes) with your extracted nucleic acids. Set up your qPCR reactions in reaction tubes or plates, ensuring proper labeling.
4. Thermal Cycling: Place the reaction tubes or plates into the PCR machine (qPCR instrument). Set the thermal cycling conditions as specified in the kit’s instructions. This includes denaturation, annealing, and extension temperatures, as well as the number of cycles.
5. Run the qPCR: Start the qPCR machine to run the amplification and detection process. The machine will cycle through temperature changes, monitoring the accumulation of fluorescent signals in real-time.

                             

6. Data Analysis: After the qPCR run is complete, analyze the data using the software provided with your qPCR machine. Interpret the results based on the kit’s guidelines. Positive and negative controls can help verify the accuracy of the assay.
7. Interpretation and Reporting: Determine the presence or absence of the targeted pathogens in your samples based on the amplification curves and cycle threshold (Ct) values. Document and report the results according to your laboratory’s standard operating procedures.
8. Quality Control: Perform quality control measures to ensure the reliability of your results, such as confirming the absence of contamination and assessing the efficiency of the qPCR assay.
9. Safety and Waste Disposal: Follow safety precautions when working with potentially infectious materials. Dispose of biohazardous waste and used materials according to local regulations.

Always remember to carefully follow the kit’s instructions and consult with a qualified laboratory professional if you are unsure about any step in the process. Accuracy and proper handling are crucial when using such diagnostic kits in a clinical or research setting.

What is the Most Common and Effective qPCR Panel Detection Kit for Sepsis?

Introducing the CHAINFOR™ Sepsis Id-30 Genotyping qPCR Kits, a breakthrough in sepsis diagnostics. These innovative kits are designed for in vitro qualitative diagnosis, specifically targeting Sepsis infection nucleic acids found in whole blood and positive blood culture samples. What sets these kits apart is their unparalleled efficiency and precision. With a ready-to-use format, you can simplify your workflow and save valuable time. The capability to detect 30 different targets in just 8 tube reactions ensures comprehensive pathogen-based genotyping, while the utilization of multiple detection channels, including FAM, Texas Red, ROX, Cy5, and HEX, guarantees accuracy. Our multiplex reaction-based procedures streamline the process without compromising results.

Moreover, with sensitivity and specificity both exceeding 97%, you can trust in the reliability of your findings. Say goodbye to lengthy waiting times; these kits provide rapid results in just 60 minutes. Whether you’re dealing with fungal or bacterial agents or assessing antibiotic resistance, our kits have you covered. CHAINFOR™ Sepsis Id-30 Genotyping qPCR Kits offer a simple, rapid, and highly sensitive solution to sepsis diagnosis, ensuring you can make informed decisions promptly.

 

REFERENCES

[1] Rudd KE, Johnson SC, Agesa KM, Shackelford KA, Tsoi D, Kievlan DR, et al. Global, regional, and national sepsis incidence and mortality, 1990-2017: analysis for the Global Burden of Disease Study. Lancet (London, England). 2020;395(10219):200-11.

[2] Huang, M., Cai, S., & Su, J. (2019). The pathogenesis of sepsis and potential therapeutic targets. International journal of molecular sciences, 20(21), 5376.

[3] Gotts JE, Matthay MA. Sepsis: pathophysiology and clinical management. British Medical Journal 2016.