scholarly journals Simultaneous detection of white spot syndrome virus (WSSV) and Taura syndrome virus (TSV) by multiplex reverse transcription-polymerase chain reaction (RT-PCR) in Pacific white shrimp Penaeus vannamei

2002 ◽  
Vol 50 ◽  
pp. 9-12 ◽  
Author(s):  
JM Tsai ◽  
LJ Shiau ◽  
HH Lee ◽  
PWY Chan ◽  
CY Lin
Keyword(s):  
Polymerase Chain Reaction ◽  
White Spot Syndrome Virus ◽  
Simultaneous Detection ◽  
Pacific White Shrimp ◽  
White Shrimp ◽  
Penaeus Vannamei ◽  
Rt Pcr ◽  
Taura Syndrome Virus ◽  
Chain Reaction ◽  
Polymerase Chain

scholarly journals Virus Detection of Pacific White Shrimp (Litopenaeus vannamei) at Fish Quarantine Center, Quality Control, and Security of Fishery Product in Surabaya I

2019 ◽  
Vol 4 (2) ◽  
Author(s):  
Aulia Azizah MS
Keyword(s):  
Polymerase Chain Reaction ◽  
Litopenaeus Vannamei ◽  
White Spot Syndrome Virus ◽  
Virus Detection ◽  
Pacific White Shrimp ◽  
White Shrimp ◽  
Chain Reaction ◽  
Pcr Polymerase Chain Reaction ◽  
Infectious Myonecrosis Virus ◽  
Polymerase Chain

Udang vaname (Litopenaeus vannamei) merupakan salah satu komoditas perikanan unggulan yang bernilai ekonomis penting dan banyak diminati oleh konsumen di pasaran. Udang vaname memiliki beberapa keunggulan yaitu pertumbuhan yang cepat, mampu beradaptasi pada kisaran salinitas yang tinggi, dan dapat dipelihara dengan sistem super intensif, namun beberapa tahun terakhir total produksi udang di Indonesia mengalami penurunan. Tahun 2012 total produksi udang menururn dari 1.900 ton menjadi 1.025 ton, virus diduga menjadi patogen yang memicu penyakit pada udang dan menyebabkan mortalitas yang tinggi. Virus yang sering menyerang budidaya udang vaname antara lain Taura Syndrome Virus (TSV), Infectious Myonecrosis Virus (IMNV), dan White Spot Syndrome Virus (WSSV). Tujuan penelitian ini yaitu untuk mendeteksi virus pada udang vaname dan mengetahui tingkat prevalensi virus yang menyerang udang vaname. Penelitian dilaksanakan pada bulan Desember 2018 – Januari 2019 di Balai Karantina Ikan, Pengendalian Mutu, dan Keamanan Hasil Perikanan Surabaya I. Materi yang digunakan pada penelitian yaitu 37 sampel post larva. Sampel diekstraksi menggunakan Silica Extraction Kit  kemudian dideteksi secara molekuler menggunakan PCR (Polymerase Chain Reaction). Penelitian ini bersifat observatif, selanjutnya data yang diperoleh dianalisis secara deskriptif. Hasil penelitian menunjukkan bahwa terdapat 3 sampel positif IMNV dan 2 sampel positif WSSV dengan nilai prevalensi sebesar 8,10% dan 5,40%. Tinggi rendahya nilai prevalensi dipengaruhi oleh tingkat penyebaran dan kondisi lingkungan yang kurang sesuai. Kata Kunci : PCR (Polymerase Chain Reaction), Prevalensi, Udang vaname, Virus.

scholarly journals Pemantauan Virus Dengan Metode PCR (Polymerase Chain Reaction) Di Pantai Utara Jawa Timur [Monitoring Virus By PCR Method (Polymerase Chain Reaction) In North Coast, East Java]

2019 ◽  
Vol 4 (1) ◽  
pp. 65
Author(s):  
Hari Suprapto, Yulia Kartika
Keyword(s):  
Water Quality ◽  
Polymerase Chain Reaction ◽  
White Spot Syndrome Virus ◽  
White Spot ◽  
Viral Diseases ◽  
Northern Coast ◽  
Taura Syndrome Virus ◽  
Chain Reaction ◽  
Yellow Head Virus ◽  
Polymerase Chain

Abstract The disease most dangerous for the cultivation activity is virus. Viruses are organisms subseluler that contain only nucleic acid (RNA or DNA) as genetic material. Koi Herpes Virus is one type of virus that causes mortality in cultured Cyprinids. KHV disease in Indonesia started in Blitar, East Java on March 2002 because the entry of imported koi fish that carry the virus KHV, while mortality prosentase could reach 80% - 85%, which causes loss of about 5 billion rupiah. In addition of KHV, there are several types of viral diseases in shrimp is White Spot Syndrome Virus (WSSV), Taura Syndrome Virus (TSV), dan Yellow Head Virus (YHV). Disease can cause losses in farming activities, such as WSSV. WSSV is an endemic disease since 1995. disease WSSV is exotic viral disease that attacks the shrimp monodon in 1998/1999 has resulted in decreased production of very large, so the Indonesian shrimp exports down 33,000 tons. Treatment of viral diseases is difficult because the virus resistant to certain antibiotics and chemical compounds. Therefore, prevention needs to be done, one through the monitoring activities conducted on the northern coast of East Java. The method implemented is monitoring in location and identification of viruses by PCR (Polymerase Chain Reaction). Monitoring in location includes water quality measurements and sampling. Identification of virus carried by IQ 2000TM. The identification procedure includes extraction, amplification and electrophoresis. Regional monitoring conducted on the northern coast of East Java includes Gresik, Lamongan, Tuban, Bangkalan, Sampang, Pamekasan, and Sumenep. Water quality at locations quite well. Results activities of monitoring on the northern coast of East Java is disease White Spot Syndrome Virus (WSSV) was found positive in several locations: Gresik, Lamongan and Tuban, while the virus Taura Syndrome Virus (TSV) and Yellow Head Virus (YHV) was not found at all locations . In tilapia, disease Koi Herpes Virus (KHV) was found positive in Tuban.

A Multiplex Reverse Transcription Polymerase Chain Reaction Method for the Detection of Foodborne Viruses†

1999 ◽  
Vol 62 (10) ◽  
pp. 1210-1214 ◽  
Author(s):  
SORAYA I. ROSENFIELD ◽  
LEE-ANN JAYKUS
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcription ◽  
Hepatitis A Virus ◽  
Simultaneous Detection ◽  
Polymerase Chain Reaction Method ◽  
Human Enterovirus ◽  
Norwalk Virus ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain

A multiplex reverse transcription polymerase chain reaction (RT-PCR) method was developed for the simultaneous detection of the human enteroviruses, hepatitis A virus (HAV) and Norwalk virus (NV). Poliovirus type 1 (PV1) was chosen as a model for the human enterovirus group. Three different sets of primers were used to produce three size-specific amplicons of 435 bp, 270 bp, and 192 bp for PV1, NV, and HAV, respectively. RT-PCR products were separated by agarose gel electrophoresis, and amplicon identity was confirmed by Southern transfer followed by DNA hybridization using nonradio-active, digoxigenin-labeled internal probes. When tested on mixed, purified virus suspensions, the multiplex method achieved detection limits of ≤1 infectious unit (PV1 and HAV) or RT-PCR-amplifiable unit (NV) for all viruses. With further streamlining efforts such as single tube amplification and liquid hybridization, multiplex PCR offers advantages over cell culture methodology and monoplex PCR because it allows for rapid and cost-effective detection of several human enteric viruses in a single reaction tube.

scholarly journals Development of multiplex reverse transcription polymerase chain reaction (RT-PCR) for simultaneous detection of matrix, haemagglutinin and neuraminidase genes of H5N1 avian influenza virus

2012 ◽  
Vol 28 (2) ◽  
pp. 55-59 ◽  
Author(s):  
R. Mojumder ◽  
E. H. Chowdhury ◽  
R Parvin ◽  
J. A. Begum ◽  
M. Giasuddin ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Influenza Virus ◽  
Avian Influenza ◽  
Matrix Protein ◽  
Simultaneous Detection ◽  
High Fidelity ◽  
Rt Pcr ◽  
Chain Reaction ◽  
One Step ◽  
Polymerase Chain

Influenza A virus, subtype H5N1 causes a fatal disease in domestic poultry and could spread directly from poultry to humans. The aim of this study was to develop a multiplex reverse transcription polymerase chain reaction (mRT-PCR) for simultaneous detection of Type A influenza virus-specific matrix protein (M) gene as well as H5 and N1 genes of highly pathogenic avian influenza (HPAI) viruses. Finnzymes Phusion-Flash High- Fidelity PCR Master Mix (Finnzymes Oy, Finland) and Qiagen one-step RT-PCR enzyme mix (Qiagen, Germany) were used in a one-step RT-PCR. RNA was extracted from two known positive samples using Qiagen RNA extraction kit. RT-PCR was carried out with a mixture of primers specific for the Type A influenza virus matrix protein (M), and H5 and N1 genes of H5N1 HPAI viruses in a single reaction system. The mRT-PCR cDNA products were visualized by gel electrophoresis. The mRT-PCR yielded fragments of 245 bp for M, 545 bp for H5 and 343 bp for N1 genes of HPAI virus, which were clearly distinguishable. The mRT-PCR using the Finnzymes Phusion-Flash High-Fidelity PCR Master Mix (Finnzymes Oy, Finland) with Qiagen one-step RT-PCR Enzyme Mix (Qiagen, Germany) required only one hour and 20 minutes. (Bangl. vet. 2011. Vol. 28, No. 2, 55 – 59)DOI: http://dx.doi.org/10.3329/bvet.v28i2.10653

scholarly journals A One-Step Reverse Transcription-Polymerase Chain Reaction System for the Simultaneous Detection and Identification of Multiple Tospovirus Infections

2005 ◽  
Vol 95 (2) ◽  
pp. 166-171 ◽  
Author(s):  
Hiroyuki Uga ◽  
Shinya Tsuda
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcription ◽  
Simultaneous Detection ◽  
Yellow Spot ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Spot Virus ◽  
Detection And Identification ◽  
One Step ◽  
Polymerase Chain

A one-step reverse transcription-polymerase chain reaction (RT-PCR) method has been developed for the simultaneous detection and identification of multiple tospoviruses that infect plants. The RT-PCR system is composed of six primers in a single tube: a universal degenerate primer and five virus species-specific primers. Amplifications resulted in an 848-bp PCR product for Watermelon silver mottle virus, 709-bp for Tomato spotted wilt virus, 589-bp for Impatiens necrotic spot virus, 511-bp for Melon yellow spot virus, and a 459-bp amplicon for Iris yellow spot virus. This system enables the simultaneous detection of at least three types of tospovirus infections, in addition to their species identities, from five possible tospoviruses studied, on the basis of their S RNA combinations. This multiplex RT-PCR system was applied to the detection of tospovirus in ornamental crops cultivated in fields and shows potential for epidemiological studies.

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