Perbandingan Metode PCR Konvensional Dengan Metode PCR Portable Kit Untuk Deteksi WSSV Pada Udang Vannamei.

Udang vannamei merupakan komoditi yang mendominasi pertambakan dipesisir wilayah Kalimantan Barat. Usaha budidaya tersebut tumbuh pesat dengan resiko penyebaran penyakit yang juga tinggi. Serangan penyakit yang paling umum dan sering ditemukan pada budidaya udang vannamei adalah WSSV ( White spot Syndrome Virus). Identifikasi penyakit virus WSSV di SKIPM Pontianak dilakukan dengan dua metode yang berbeda yaitu metode PCR Konvensional dan Portable Kit PCR. Metode PCR konvensional merupakan salah satu alternatif untuk deteksi penyakit virus yang cukup akurat dan relatif lebih murah, jika dibandingkan dengan metode lain yang sedang berkembang saat ini seperti metode PCR Portable Kit yang proses deteksi lebih singkat karena tidak memerlukan tahap elektroforesis. Melihat dari kelebihan dan kekurangan masing- masing metode tersebut, sehingga menimbulkan alasan untuk mengetahui lebih lanjut tentang presisi dan akurasi sensitivitas dari kedua metode tersebut dalam mendeteksi infeksi virus WSSV pada udang vannamei. Pengamatan dilakukan pada variabel plasmid kontrol (+) masing – masing kit uji, plasmid DNA sampel dan plasmid DNA virus WSSV dengan indikator hasil pemeriksaan positif, negatif, terdeteksi maupun tidak terdeteksi. Setiap hasil pemeriksaan variabel pengamatan akan diberi nilai pembobotan tertentu, untuk mengetahui presisi dan akurasi sensitivitas dari kedua metode PCR dalam mendeteksi infeksi virus WSSV pada udang vannamei.

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Hijacking of Host Calreticulin is Required for the White Spot Syndrome Virus Replication Cycle.

Journal of Virology ◽

10.1128/jvi.01014-14 ◽

2014 ◽

pp. JVI.01014-14 ◽

Cited By ~ 5

Author(s):

Apiruck Watthanasurorot ◽

Enen Guo ◽

Sirinit Tharntada ◽

Chu-Fang Lo ◽

Kenneth Söderhäll ◽

...

Keyword(s):

Endoplasmic Reticulum ◽

White Spot Syndrome Virus ◽

Replication Cycle ◽

Host Protein ◽

Host Cells ◽

White Spot ◽

Structural Proteins ◽

Dna Virus ◽

White Spot Syndrome ◽

Viral Structural Proteins

We have previously shown that multifunctional calreticulin (CRT), which resides in the endoplasmic reticulum (ER) and is involved in ER-associated protein processing, responds to infection with white spot syndrome virus (WSSV) by increasing mRNA and protein expression and by forming a complex with gC1qR and thereby delaying apoptosis. Here, we show that CRT can directly interact with WSSV structural proteins, including VP15 and VP28, during an early stage of virus infection. The binding of VP28 with CRT does not promote WSSV entry, and CRT-VP15 interaction was detected in the viral genome in virally infected host cells, and thus may have an effect on WSSV replication. Moreover, CRT was detected in the viral envelope of purified WSSV virions. CRT was also found to be of high importance for proper oligomerization of the viral structural proteins VP26 and VP28, and when CRT glycosylation was blocked with tunicamycin, a significant decrease in both viral replication and assembly was detected. Together, these findings suggest that CRT confers several advantages to WSSV from the initial steps of WSSV infection, to the assembly of virions. Therefore, CRT is required as a “vital factor” and is hijacked by WSSV for its replication cycle.ImportanceWhite spot syndrome virus (WSSV) is a double-stranded DNA virus and the cause of a serious disease in a wide range of crustaceans that often leads to high mortality rates. We have previously shown that the protein calreticulin (CRT), which resides in the endoplasmic reticulum (ER) of the cell, is important in the host response to the virus. In this report, we show that the virus uses this host protein to enter the cell and to make the host produce new viral structural proteins. Through its interaction with two viral proteins, the virus “hijacks” host calreticulin and uses it for its own needs. These findings provide new insight into the interaction between a large DNA virus and the host protein CRT, and may help in understanding the viral infection process in general.

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