scholarly journals Reverse Transcriptase PCR Detection of Astrovirus, Hepatitis A Virus, and Poliovirus in Experimentally Contaminated Mussels: Comparison of Several Extraction and Concentration Methods

1998 ◽  
Vol 64 (8) ◽  
pp. 3118-3122 ◽  
Author(s):  
Ousmane Traore ◽  
Charlotte Arnal ◽  
Berengere Mignotte ◽  
Armand Maul ◽  
Henri Laveran ◽  
...  
Keyword(s):  
Reverse Transcriptase ◽  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Extraction Methods ◽  
Borate Buffer ◽  
Rt Pcr ◽  
Peg 6000 ◽  
Reverse Transcriptase Pcr ◽  
Glycine Solution ◽  
Concentration Methods

ABSTRACT Four methods of extraction and three methods of concentration of three enteric viruses from mussels were comparatively evaluated by reverse transcriptase PCR (RT-PCR). Shellfish were experimentally contaminated by immersion in seawater seeded with astrovirus, hepatitis A virus, or poliovirus. Sixty-gram samples of mussel tissues were processed by using borate buffer, glycine solution, saline beef, and saline beef-Freon extraction methods. The viruses were concentrated by precipitation with polyethylene glycol 6000 (PEG 6000) or PEG 8000 or by organic flocculation. RT-PCR was performed with RNA extracts from crude shellfish extracts and concentrates with and without Sephadex LH20 filtration. The glycine solution and borate buffer extraction methods resulted in significantly more RT-PCR-positive samples than the saline beef extraction method. We assessed the efficiency of 20 combinations of extraction and concentration methods. The borate buffer-organic flocculation, borate buffer-PEG 6000, and glycine solution-PEG 6000 combinations gave RT-PCR-positive results for all 27 samples analyzed for the three viruses. Detoxification of the samples by Sephadex LH20 filtration significantly decreased the efficiency of RT-PCR virus detection.

Application of Concanavalin A–Linked Magnetic Beads for the Detection of Hepatitis A Virus

2018 ◽  
Vol 81 (12) ◽  
pp. 1997-2002
Author(s):  
SANG-MU KO ◽  
SE-YOUNG CHO ◽  
MYUNG-JOO OH ◽  
JOSEPH KWON ◽  
BIPIN VAIDYA ◽  
...  
Keyword(s):  
Reverse Transcriptase ◽  
Binding Affinity ◽  
Concanavalin A ◽  
Hepatitis A ◽  
Magnetic Beads ◽  
Hepatitis A Virus ◽  
Immunomagnetic Separation ◽  
Carbohydrate Binding ◽  
Con A ◽  
Reverse Transcriptase Pcr

ABSTRACT Prompt and inexpensive detection of hepatitis A virus (HAV) is essential to control acute hepatitis outbreaks associated with the consumption of contaminated raw or minimally processed food. In this study, various carbohydrate-binding lectins, including concanavalin A (Con A), wheat germ agglutinin, and soybean agglutinin, were compared for their binding affinity to HAV. Con A, which showed significantly higher binding affinity than other lectins, was used to develop an alternative and affordable method to conventional antibody-linked immunomagnetic separation prior to detection of HAV using reverse transcriptase PCR. This method, Con A–linked immunomagnetic separation combined with reverse transcriptase PCR, can detect HAV at a dilution concentration of 10−4 of the virus stock (titer: 104 median tissue culture infective dose per mL), indicating that Con A could be a promising candidate for concentrating HAV.

DETECTION AND DIFFERENTIATION OF THE VACCINE STRAIN AND FIELD ISOLATES OF DUCK HEPATITIS A VIRUS TYPE 1 USING REAL-TIME RT-PCR AND HIGH RESOLUTION MELTING ASSAYS

2015 ◽  
Vol 41 (04) ◽  
pp. 229-235
Author(s):  
Kuang-Po Li ◽  
Shan-Chia Ou ◽  
Jui-Hung Shien ◽  
Poa-Chun Chang
Keyword(s):  
High Resolution ◽  
Real Time ◽  
Virus Type ◽  
Vaccine Strain ◽  
Hepatitis A ◽  
High Resolution Melting ◽  
Hepatitis A Virus ◽  
Rt Pcr ◽  
Duck Hepatitis

Duck hepatitis A virus type 1 (DHAV-1) infection is a highly contagious and fatal disease of young ducklings. A live attenuated vaccine strain designated as 5886 has been used in Taiwan for the control of DHAV-1. Although several molecular biological methods are reported for diagnosis of DHAV-1 infection, none of them is able to discriminate between the vaccine strain and field viruses of DHAV-1. In the present study, a real-time reverse transcriptase polymerase chain reaction (RT-PCR) and high resolution melting (HRM) assay was developed for rapid detection and differentiation between the vaccine strain and field viruses of DHAV-1. This assay is highly specific for DHAV-1 and the detection limit is about 100 copies of the viral RNA. Experiments using fecal samples collected from ducklings experimentally infected with DHAV-1 showed that DHAV-1 could be detected in fecal samples as early as 6 h post-infection. In summary, a real-time RT-PCR and HRM assay is developed in this study and this assay could be valuable for diagnosis and surveillance of DHAV-1 infection in the field.

Comparison of different concentration methods for the detection of hepatitis A virus and calicivirus from bottled natural mineral waters

2010 ◽  
Vol 165 (1) ◽  
pp. 57-63 ◽  
Author(s):  
S. Di Pasquale ◽  
M. Paniconi ◽  
B. Auricchio ◽  
L. Orefice ◽  
A.C. Schultz ◽  
...  
Keyword(s):  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Mineral Waters ◽  
Natural Mineral ◽  
Concentration Methods

Development of a One-Step Duplex RT-PCR Method for the Simultaneous Detection of VP3/VP1 and VP1/P2B Regions of the Hepatitis A Virus

2016 ◽  
Vol 26 (8) ◽  
pp. 1398-1403 ◽  
Author(s):  
Mi-Ju Kim ◽  
Shin-Young Lee ◽  
Hyun-Joong Kim ◽  
Jeong Su Lee ◽  
In Sun Joo ◽  
...  
Keyword(s):  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Simultaneous Detection ◽  
Rt Pcr ◽  
One Step ◽  
Pcr Method

scholarly journals First Genome Sequence of Newcastle Disease Virus of Genotype VIIi from Jordan

2018 ◽  
Vol 7 (23) ◽  
Author(s):  
Mustafa Ababneh ◽  
Helena L. Ferreira ◽  
Mohammad Khalifeh ◽  
David L. Suarez ◽  
Claudio L. Afonso
Keyword(s):  
Reverse Transcriptase ◽  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Genome Sequence ◽  
Illumina Sequencing ◽  
Newcastle Disease ◽  
Rt Pcr ◽  
Total Rna ◽  
Reverse Transcriptase Pcr ◽  
Chicken Spleen

Newcastle disease virus (NDV) was detected by reverse transcriptase PCR (RT-PCR) from total RNA isolated from a chicken spleen of a backyard flock in Jordan. The complete coding genome sequence of NDV/chicken/Jordan/J11-spleen/2018 was obtained with MiSeq (Illumina) sequencing.

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