H1N1 PCR test for Swine Flu in Hyderabad - Cost and Home Collection

The H1N1 test process includes the detection of H1N1 virus. The diagnosis of h1n1 flu (influenza A) can be difficult because the virus's symptoms (H1N1) are similar to those of other respiratory viruses, such as rhinoviruses, adenoviruses, parainfluenza, and respiratory syncytial virus. The incubation period for Influenza is 2 to 3 days, and a viral sample from a patient's nose and throat swabs will be collected.

• Within 2 to 3 days from the day of infection, 
• Before the administration or use of antiviral drugs and 
• After the onset of H1N1 symptoms. 

The H1N1 swab test procedure includes the insertion of a dry swab into the nostril and back of the nasopharynx (upper part of the throat behind the nose), where the technician leaves the swab for a few seconds. Slowly remove the swab while slightly rotating. Another new swab will be used for the adjacent nostril, and the same process will be repeated. The tip of the swab will be placed into a Viral Transport Medium (VTM) filled vial by breaking the applicator stick.

A nasopharyngeal swab, nasal wash, or nasal aspirate are preferred respiratory samples to use for influenza H1N1 test. If the above samples are unavailable, a nasal swab and/or oropharyngeal swab will be acceptable. In intubated patients, an endotracheal aspirate will be collected.



These samples will be preserved in a sterile viral transport medium, kept under cold temperature (on ice, in cold packs, or the refrigerator, preferably at 4°C), and sent for further analysis to the specialised laboratories within 48 hours of transportation.

The influenza virus (H1N1) first appeared in April 2009 in Mexico and quickly spread over the world. On the 11th of June, 2009, the World Health Organization (WHO) proclaimed a global pandemic of phase 6 level, which lasted until the 10th of August, 2010 and resulted in multiple deaths worldwide. Classical swine H1N1 was originally identified in pigs in the United States in 1930, following the 1918 pandemic of the same name.

There are many possible detection tests available, each with its own level of specificity. The detection techniques for influenza A(H1N1) can be classified into the following categories:

Clinical Diagnosis (CD)

1. Virus culture
2. Immunofluorescence assays

Surveillance Method (SM)

1. Rapid influenza detection tests
2. PCR-based detection methods
3. Non PCR based RNA specific detection methods
4. Biosensors

The PCR test for H1N1 is a standard laboratory method for detecting the H1N1 virus by multiplying a segment of DNA to several times.



Polymerase is an enzyme responsible for the formation of DNA polymers. It only detects the presence of DNA. The chain reaction is a chemical reaction that progresses in an exponential form. The polymerised chain reaction is a laboratory technique used to identify the type and properties of the virus by amplifying its DNA.

In general, there will be a conversion of DNA to messenger RNA (mRNA) through a process known as transcription. In the reverse transcriptase polymerase chain reaction (RT-PCR), as the polymerase detects only DNA molecules, the RNA of the virus will be converted into DNA, a process called reverse transcription (RT). With the help of the PCR technique, the single viral DNA will be amplified and will produce several copies (polymers) of viral DNA, leading to the accurate detection of viral presence. 

A sample will be collected from the areas such as the nose and throat of the patient, and the obtained sample will be treated with certain chemicals to remove proteins, fats and extract only the viral RNA.

The viral RNA will be converted to DNA by a reverse transcription mechanism with the help of certain enzymes. 

Taq probe, DNA polymerase, reverse transcriptase enzyme, primers, and nucleotides are the essential components of a polymerase chain reaction (PCR). DNA synthesis involves assembling these components in a tube with the necessary cofactors for the enzyme and then subjecting the mixture to repeated heating and cooling cycles.

The mixture along with extracted RNA will be placed in an RT-PCR machine; the machine heats and cools the mixture to trigger specific chemical reactions and makes several copies of viral DNA in an exponential form.

Each RT-PCR cycle follows the basic steps after the formation of DNA from viral RNA (reverse transcription) such as 

• Denaturation 
• Annealing 
• Extension
  

The obtained DNA copies go through the same procedure, and each cycle produces an exponential number of DNA copies. The standard PCR process involves 35 cycles and produces 35 billion new DNA copies, as well as an equivalent quantity of fluorescence. This fluorescence will be recorded as digital data using Real-time PCR.