Establishment of RT-PCR and Real Time RT-PCR assays for diagnosis of Nipah virus encephalitis
Keywords:
Nipah virus, RT-PCR, Real Time RT-PCRAbstract
Nipah virus (NiV) still remains a public health problem in Asia after its first outbreak in Malaysia during 1998-1999. NiV-associated encephalitis is known to cause a high mortality rate because of no specific treatment. Laboratory investigation for NiV infection, therefore, plays an important role on care, prevention and control. To strengthen laboratory capacity and capability on NiV diagnosis, we have established detection methods for NiV-associated encephalitis diagnosis which are Reverse transcription–polymerase chain reaction (RT-PCR) and Real time RT-PCR. The techniques aim to detect N gene of Nipah virus. Our study demonstrated that the limit of detection of RT-PCR and Real time RT-PCR assays was 104 copies/reaction and 102 copies/reaction, respectively. The assays were shown to be specific for NiV only. Determination of NiV in cerebrospinal fluid (CSF) from 30 patients with encephalitic symptoms found negative for N gene, suggesting that at present there is no Nipah infection in Thailand.
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