Development of Loop-mediated Isothermal Amplification for detection of Fowl Adenovirus serotype 2 causing Inclusion body Hepatitis
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Abstract
Inclusion Body Hepatitis (IBH) is a disease with a high mortality rate in chicks, causing significant devastation to the poultry production industry worldwide. Thailand first encountered an outbreak of IBH in 2007, caused by the fowl adenovirus serotype 2 (FAdV-2). To detect this virus, the study developed a loop-mediated isothermal amplification (LAMP) method using four primers specific to the hexon gene. The optimal results were obtained at a temperature of 63 degrees Celsius for 60 minutes. These results can be interpreted with the naked eye due to the addition of SYBR Green I in reaction mixture, and no reactions were detected with the genetic material of others viruses such as chicken anemia virus (CAV), Newcastle disease virus (NDV), duck plaque virus (DPV), infectious bronchitis virus (IBV), and egg drop syndrome virus (EDSV). The LAMP technique could detect virus levels as low as 10 copies/µl, making it 1000 times more sensitive than polymerase chain reaction (PCR) techniques. This study concludes that the LAMP technique is suitable for diagnosing IBH caused by the FAdV-2 virus. It is a simple, convenient, and fast method that does not require expensive equipment for testing and interpreting results. The technique has high specificity and sensitivity, making it ideal for fieldwork.
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