TY  - JOUR
T1  - Single-Step Real-Time Reverse Transcription-Polymerase Chain Reaction for Simultaneous Detection of H5N1 and H5N8 Highly Pathogenic Avian Influenza Viruses
AU - Park, Choi-Kyu AU - Kim, Eun-Mi 
JO  - Journal of Animal and Veterinary Advances
VL  - 14
IS  - 6
SP  - 161
EP  - 166
PY  - 2015
DA  - 2001/08/19
SN  - 1680-5593
DO  - javaa.2015.161.166
UR  - https://makhillpublications.co/view-article.php?doi=javaa.2015.161.166
KW  - Highly pathogenic avian influenza virus
KW  -real-time RT-PCR
KW  -H5N1
KW  -H5N8
KW  -detection
AB  - For efficient monitoring of Highly Pathogenic Avian Influenza Viruses (HPAIVs) in countries where various subtype H5 HPAIVs co-circulate in avian populations, a Simultaneous Differential Detection Method for major and minor subtypes of HPAIVs is needed. In this study, we developed a single-step Real-time Reverse Transcription-Polymerase Chain Reaction (sRRT-PCR) assay for the simultaneous detection of <I>HPAIVM</I>, <I>H5</I>, <I>N1</I> and <I>N8</I> genes. The <I>M</I> gene primer set in the assay detected all subtypes of AIV including representative H5N1 and H5N8 strains from Korea as reported in earlier studies and the <I>H5</I>, <I>N1</I> and <I>N8</I> gene primer sets were able to specifically detect H5, N1 and N8 subtypes of HPAIV, respectively. The detection limits for the <I>M</I>, <I>H5</I>, and <I>N1</I> or <I>N8</I> genes were 0.6, 0.7 and 0.9 EID<SUB>50</SUB> for H5N1 HPAIV and 1.0, 0.8 and 1.3 EID<SUB>50</SUB> for H5N8 HPAIV, respectively. The assay was sufficiently sensitive for monitoring and surveillance of HPAIVs and can be used in the differential diagnosis of H5N1, H5N8 and other subtypes of AIV in countries where subtype H5 HPAIV outbreaks are occurring.
ER  -