scholarly journals Polymerase Chain Reaction (PCR) Amplification of DNA of Red Sea Bream Iridovirus (RSIV)

1998 ◽  
Vol 33 (1) ◽  
pp. 17-23 ◽  
Author(s):  
Jun Kurita ◽  
Kazuhiro Nakajima ◽  
Ikuo Hirono ◽  
Takashi Aoki
Keyword(s):  
Polymerase Chain Reaction ◽  
Red Sea ◽  
Pcr Amplification ◽  
Sea Bream ◽  
Red Sea Bream ◽  
Chain Reaction ◽  
Polymerase Chain

scholarly journals Rapid diagnosis of red sea bream iridovirus infection using the polymerase chain reaction

1998 ◽  
Vol 32 ◽  
pp. 87-90 ◽  
Author(s):  
S Oshima ◽  
JI Hata ◽  
N Hirasawa ◽  
T Ohtaka ◽  
I Hirono ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Red Sea ◽  
Rapid Diagnosis ◽  
Sea Bream ◽  
Red Sea Bream ◽  
Chain Reaction ◽  
Polymerase Chain

scholarly journals Deteksi Spesies Megalocytivirus pada Budidaya Ikan Laut di Kepulauan Riau

2019 ◽  
Vol 13 (3) ◽  
pp. 265-274
Author(s):  
Achmad Bahtiar Rifai ◽  
Anak Agung Gede ◽  
Lucky Putra P Kusuma ◽  
Yani Maulani
Keyword(s):  
Polymerase Chain Reaction ◽  
Capsid Protein ◽  
Red Sea ◽  
Major Capsid Protein ◽  
Sea Bream ◽  
Red Sea Bream ◽  
Necrosis Virus ◽  
Chain Reaction ◽  
Polymerase Chain

Penyakit yang dapat menyebabkan kerugian ekonomi yang sangat tinggi adalah virus dan salah satunya adalah Megalocytivirus. Megalocytivirus mempunyai tiga spesies yaitu Infectious Spleen and Kidney Necrosis Virus (ISKNV), Red Sea Bream Iridovirus (RSIV), dan Turbot Redy Body Iridovirus (TRBIV). Gejala klinis pada ikan yang terinfeksi Megalocytivirus pada umumnya terlihat dari perubahan tingkah laku yaitu nafsu makan berkurang, berenang lemah, dan malas bergerak. Tubuh ikan yang terinfeksi terlihat lebih gelap baik pada permukaan tubuh sirip, dan bagian ekor. Metode uji yang digunakan pada penelitian ini adalah pengambilan sampel, pengujian histopatologi, deteksi spesies Megalocytivirus dengan polymerase chain reaction (PCR). Sampel ikan yang menunjukan gejala klinis diambil dari beberapa kabupaten/kota yang ada di kepulauan Riau antara lain Batam, Tanjungpinang, Bintan, Tanjung Balai Karimun dan Natuna. Sampel ikan adalah ikan kakap dan kerapu dari keramba jaring apung dan hatchery. Tujuan dari penelitian ini adalah mendeteksi spesies Megalocytivirus pada budidaya ikan laut yang ada di Kepulauan Riau. Pada uji histopatologi ditemukan lesio berupa badan inklusi, nekrosa, melano makrofag center pada hati, ginjal dan limpa. Badan inklusi dan nekrosa yang menyebar pada organ tersebut merupakan karakteristik Megalocytivirus. Hasil identifikasi PCR dengan menggunakan gen Major Capsid Protein pada panjang amplikon 777 bp berhasil mendeteksi Megalocytivirus. deteksi spesies Megalocytivirus dibeberapa Kabupaten/Kota di Kepulauan Riau menunjukan semua sampel ikan air laut terinfeksi ISKNV, sedangkan untuk spesies RSIV dan TRBIV tidak berhasil dideteksi pada penelitian ini. Infeksi Mengalocytivirus sudah menyebar pada budidaya ikan laut diseluruh kabupaten/Kota di Propinsi Kepulauan Riau. Spesies Megalocytivirus yang terdeteksi di Kepulauan Riau adalah ISKNV.

scholarly journals Evaluation of a novel TaqMan probe-based real-time polymerase chain reaction (PCR) assay for detection and quantitation of red sea bream iridovirus

2021 ◽  
Vol 24 (11) ◽  
pp. 351-359
Author(s):  
Guk Hyun Kim ◽  
Min Jae Kim ◽  
Hee Ju Choi ◽  
Min Ji Koo ◽  
Min Jeong Kim ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Red Sea ◽  
Sea Bream ◽  
Taqman Probe ◽  
Red Sea Bream ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Polymerase Chain

scholarly journals Amplification of human beta-actin gene by the reverse transcriptase-polymerase chain reaction: implications for assessment of RNA from formalin-fixed, paraffin-embedded material.

1996 ◽  
Vol 44 (10) ◽  
pp. 1205-1207 ◽  
Author(s):  
A Dakhama ◽  
V Macek ◽  
J C Hogg ◽  
R G Hegele
Keyword(s):  
Polymerase Chain Reaction ◽  
Pcr Amplification ◽  
Housekeeping Gene ◽  
Chain Reaction ◽  
Viral Genomes ◽  
Negative Results ◽  
Beta Actin ◽  
Target Nucleic Acid ◽  
Formalin Fixed Paraffin Embedded ◽  
Polymerase Chain

The polymerase chain reaction (PCR) is a powerful method that allows enzymatic amplification of rate target nucleic acid sequences. It has been applied to the amplification of viral genomes from paraffin-embedded pathology specimens. However, interpretation of negative results requires amplification of a housekeeping gene such as beta-actin. In the present study we used specific oligonucleotide primers previously designed to amplify both the genomic DNA and the mRNA transcript from paraffin-embedded tissue. These products have predicted sizes of 250 BP and 154 BP, respectively, but our results showed that PCR amplification only (without reverse transcription) unexpectedly generated the 154-BP product. Further investigation of the nature of this product demonstrated that it originated from the amplification of DNA, not RNA. We conclude that the 154-BP product generated by these primers cannot be exclusively considered as beta-actin RNA product and should not be used to assess successful extraction of RNA, to ascertain its integrity, or to normalize for the total amount of RNA assayed by RT-PCR from paraffin-embedded tissue.

scholarly journals A Self-Contained Portable Polymerase-Chain-Reaction System Integrated with Electromagnetic Mini-Actuators for Bi-Directional Fluid Transport

2011 ◽  
Vol 27 (3) ◽  
pp. 357-364
Author(s):  
B. T. Chia ◽  
S.-A. Yang ◽  
M.-Y. Cheng ◽  
C.-W. Lin ◽  
Y.-J. Yang
Keyword(s):  
Polymerase Chain Reaction ◽  
Fluid Transport ◽  
Point Of Care ◽  
Reaction System ◽  
Thermal Characteristics ◽  
Pcr Amplification ◽  
Chain Reaction ◽  
Rapid Temperature ◽  
Small Footprint ◽  
Polymerase Chain

ABSTRACTIn this paper, the development of a portable polymerase chain reaction (PCR) device is presented. Integrating electromagnetic mini-actuators for bi-directional fluid transport, the proposed device, whose dimension is 67mm × 66mm × 25mm, can be fully operated with a 5V DC voltage. The device consists of four major parts: A disposable channel chip in which PCR mixture is manipulated and reacted, a heater chip which generates different temperature zones for PCR reaction, a linear actuator array for pumping PCR mixture, and a circuit module for controlling and driving the system. The advantages of the device include the rapid temperature responses associated with continuous-flow-type PCR devices, as well as the programmable thermal cycling associated with chamber-type PCR devices. The thermal characteristics are measured and discussed. PCR amplification is successfully performed for the 122 bp segment of MCF-7/adr cell line. Due to its small footprint, this self-contained system potentially can be employed for point-of-care (POC) applications.

scholarly journals Method Comparison of DNA Isolation and Quantification for Fish and Seafood Authenticity Determination

2018 ◽  
Vol 13 (3) ◽  
pp. 115
Author(s):  
Dwiyitno Dwiyitno ◽  
Stefan Hoffman ◽  
Koen Parmentier ◽  
Chris Van Keer
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Extraction ◽  
Dna Isolation ◽  
Blue Mussel ◽  
Pcr Amplification ◽  
Amplification Efficiency ◽  
Chain Reaction ◽  
Seafood Products ◽  
Polymerase Chain ◽  
Commercial Kit

Fish and seafood products has been commonly targeted for fraudulent activities. For that reason, authentication of fish and seafood products is important to protect consumers from fraudulent and adulteration practices, as well as to implement traceability regulation. From the viewpoint of food safety, authenticity is beneficial to protect public from serious food poisoning incidents, such as due to ingestion of toxic species. Since DNA based identification depends on the nucleic acid polymerase chain reaction (PCR), the quantity and quality/purity of DNA will contribute significantly to the species authentication. In the present study, different DNA extraction and purification methods (3 classical methods and one commercial kit) were compared to produce the better isolated DNA for PCR amplification. Additionally, different methods for the estimation of DNA concentration and purity which is essential for PCR amplification efficiency were also evaluated. The result showed that classical DNA extraction methods (based on TNES-Urea) yielded a higher amount of DNA (11.30-323.60 ng/g tissue) in comparison to commercial kit/Wizard Promega (5.70-83.45 ng/g tissue). Based on the purity of DNA extract (A260/280), classical DNA extraction method produced relatively similar on DNA quality to the commercial kit (1.79-2.12). Interestingly, all classical methods produced DNA with A260/280 ratio of more than 2.00 on the blue mussel, in contrast with commercial kit. The commercial kit also produced better quality of DNA compared to the classical methods, showing the higher efficiency in PCR amplification. NanoDrop is promising as cheap, robust and safe UV-spectrophotometer method for DNA quantification, as well as the purity evaluation.Keywords: seafood authenticity, DNA isolation, polymerase chain reaction, NanoDrop, Picogreen

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