H1N1 (Swine Flu Virus): Real-Time PCR Recommended Method for Confirmation of Infection

H1N1 is a subtype of influenza virus A. This subtype was responsible for the 2009 pandemic and responsible for 284,400 deaths worldwide. In 2012 and 2015, India also reported many cases of H1N1 including approx. 2000 deaths. H1N1 is commonly known as the swine flu virus because it's commonly found in swine (Pigs) and due to antigenic shift, the virus can make genetic changes that make it able to infect the human population living in direct contact with pigs. The H1N1 name indicates the viral protein that helps the virus to infect the cells and move from one host cell to another: Hemagglutinin (H) and Neuraminidase (N). These antigen proteins also help in classifying the Influenzas’ A subtypes, such as H1N1, H3N2, H1N2, etc.

Other Name: H1N1 Real-time PCR, H1N1 RNA qualitative PCR, Swine flu PCR

H1N1 Flu symptoms

H1N1 flu symptoms are very similar to other flu symptoms, which include:

Fever With Chills And Sweats (Not Always)
Muscle Pain
Sore Throats
Cough
Cold With Runny Nose
Eye Pain With Red And Watery Eyes
Headache
Body Aches
Tiredness
Weakness
Stomach Pain
Vomiting
Diarrhoea

After 3-6 days from the early onset of flu symptoms, most people recover on their own but some people infected with H1N1 can develop pneumonia or acute respiratory distress syndrome (ARDS) either because of viral or secondary bacterial pneumonia.

H1N1 Transmission

The H1N1 pandemic also happened because like covid, H1N1 also very easilytransmits from one person to another through respiratory droplets from coughing and sneezing. A person can also get the H1N1 virus from the contaminated surfaces.

Who Should Get Tested?

Pregnant women with Flu symptoms
A person experiences above mention flu symptoms and has preexisting chronic diseases.
If you express some of the following emergency flu symptoms: chest pain with shortness of breath, dizziness, and seizures, in children, Severe pain and trouble breathing, and change in skin colour to pale, grey, or blue.

Prevention and Treatment

The Center for Disease Control and Prevention (CDC) recommends annual flu vaccination for everyone aged 6 months or older. It's important to take measures to prevent getting the flu and limit its spread such as Wash your hands often, Cover while coughing and sneezing, avoiding touching your eyes, nose, and mouth unnecessarily, and cleaning and disinfecting surfaces you come in contact with, and avoiding coming in contact with Flu sick person.
There is no primary/ direct medication available for H1N1 treatment but secondary/indirect medication can help to get rid of the virus. Low - Mild symptoms can be treated at home with rest, oral hydration, and antipyretics medications. Severe symptoms can complicate the health condition that can be treated at the hospital by providing secondary treatment (intravenous (IV) hydration, correction of electrolyte imbalances, antibiotics to prevent bacterial concomitant and intensive care units in response to respiratory failure or sepsis, and Antiviral drugs.

H1N1 Diagnostic

The two of the widestmethods for H1N1 diagnosis are Rapid influenza A antigen diagnostic tests (RIDTs) and direct and indirect immunofluorescence tests for influenza A, but these tests are not confirmatory because of low sensitivity (10-70%) and non-specificity for detecting H1N1 influenza in clinical specimens. The real-time PCR methodis the most reliable diagnostic test for H1N1. H1N1 Real-Time PCR is the recommended method for confirmation of infection with H1N1.

H1N1 Real-Time PCR

H1N1 Real-Time PCR based on the Reverse transcription and TaqMan chemistry for detection of H1N1 virus in a patient sample.

Sample Collection:Oropharyngeal or nasopharyngeal swab specimen collected aseptically by a trained technician from the patient nose and throat. The swap is then inserted in a Viral Transport Medium (VTM) for transport and further analysis.
Sample Transport: Collected swap sample in VTM stored at room temperature (25^oC) and transported to testing molecular Lab.
DNA isolation: DNA is isolated from the VTMusing fully automated Nucleic acid extraction machines after a vigorous vortex of the VTM tube. After that isolated RNA genetic material of the virus is amplified with the help of a Real-Time PCR machine.
Real-time PCR: In Real-Time PCR, whole RNA isolated from the patient sample is 1^st converted to complementary DNA (cDNA) by a method called reverse transcription with the help of reverse transcriptase enzyme. The cDNA was then amplified for specific conserved sequences of the H1N1 nucleocapsid gene and hemagglutinin geneand detected in real-time as it was amplified with the help of fluorescent probes. The amplification of the target sequence of H1N1 causes the fluorescent probe in the test master to release the fluorophores in an exponential mannerwhich is then captured by the built-in optoelectronic sensor and displayed as an amplification curve on the screen, on a real-time basis during the test run.
Result Analysis: The doctor will analyze the result, whether the H1N1 RNA is amplified or not, which concludes the positive or Negative result for H1N1 infection. Finally, the Report is authenticated and released.

Positive / Detected H1N1 result Mean:A positive result means that H1N1 RNA is present in the blood sample. The H1N1 Real Time PCR test is semi quantitative test therefore positive result report will also mention the Cycle threshold (Ct) value. A control Ct values indicate the result is valid and test Ct (H1N1 Ct) indicates the risk of infection. The Ct value is the number of amplification cycles required for

the fluorescent signal to cross the threshold of the background fluoresce noise.The lower Ct value means a higher viral amount in the patient and the patient is likely to have an active H1N1 infection. A lower Ct value is associated with a greater risk.

Negative Result:A negative result means that H1N1RNA is not detected in the sample. This can indicate either that the person is not infected with H1N1 or that the viral load is too low to be detected by the test.

Pre-Requirement and Risk Associated with H1N1 Real-Time PCR Testing

There is no special Pre-requirement for testing however it’s better to inform the phlebotomist if you are taking any medication or have any allergy or medical condition. There is no risk involved in H1N1 testing, therefore patient does not require any special attention after testing.

H1N1 Qualitative Real-time PCR Test Price in Delhi /NCR

The price of the test varies from lab to lab and depends on Lab certification; the instrument used, and qualified technical staff and doctors.

H1N1 Qualitative Real-time PCR test at Ganesh Diagnostic and Imaging Centre

Ganesh Diagnostic and Imaging Centre is a NABL-accredited lab. NABL accreditation reflects the competency of laboratory infrastructure, technical staff, and equipment based on national and international standards.
The lab has state-of-the-art genetic material extraction to target amplification machines such asBio-Rad Real-Time PCR machine, fully closed Truelab Quattro Real-Time Quantitative PCR Machine, Genetix HT 96 Purifier Machine, APS 32 Extractor, Automated MolBio extractors, and qualified techniques to perform testing and doctor for result analysis and report authentication.
The use of fully automated machines at Ganesh Diagnostic eliminates the errors that can happen due to human intervention. All this makes sure that the patient will receive a reliable and 100% Accurate Diagnostic Report.
The Test Report in 24 hrfrom the sample collection for the H1N1 Qualitative Real-time PCR.
Free Home collection and online reportfacilities are provided by Ganesh Diagnostic so that patients can get tested from the comfort of their homes.
The Pathology services operate 24x7, 365 days, providing service to patients round the clock!
At Ganesh Diagnostic and Imaging Centre, all diagnostic test prices are very reasonable, and currently, the H1N1 Qualitative Real-time PCR Test is done at 50% OFF.