Direct sequencing of hepatitis A virus and norovirus RT-PCR products from environmentally contaminated oyster using M13-tailed primers

Background: Human rhinovirus (HRV) is still the most prevalent viral infection in humans and a significant cause of acute respiratory tract infections (ARTIs) in many communities, including military personnel undergoing basic training. Objectives: In this research, we assessed the molecular epidemiology, genotyping, and phylogenetic classification of HRVs in Iranian military trainees with respiratory infections (RI). Methods: For HRV identification and genotyping, respiratory specimens were obtained, and RT-PCR was conducted for genotyping and phylogenetic analysis of HRV utilizing primers for the 5-UTR region. Results: Among 400 Iranian military trainees (average age of 21 ± 4 years, the range of 18 - 57 years) with respiratory infections, HRV was detected in 29 patients (7%) using RT-PCR. The direct sequencing of PCR products from 10 specimens showed that the incidence of type A (n = 5, 50%) was higher than that of type B (n = 4, 40%) and type C (n = 1, 10%). There were no significant associations between HRV and respiratory and clinical symptoms, blood group, and indoor or outdoor conditions (P-value > 0.05). Conclusions: This research was the first to record HRV as a significant cause of respiratory problems among military trainees in Iran, with a frequency of 7%. The most prevalent genotype was HRV-A, which may be applicable in epidemiological and clinical studies, as well as vaccination plans.

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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

Taiwan Veterinary Journal ◽

10.1142/s1682648515500146 ◽

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.

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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

Journal of Microbiology and Biotechnology ◽

10.4014/jmb.1604.04047 ◽

2016 ◽

Vol 26 (8) ◽

pp. 1398-1403 ◽

Cited By ~ 1

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

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Detection of enteroviruses and hepatitis a virus in water by consensus primer multiplex RT-PCR

World Journal of Gastroenterology ◽

10.3748/wjg.v8.i4.699 ◽

2002 ◽

Vol 8 (4) ◽

pp. 699 ◽

Cited By ~ 14

Author(s):

Jun-Wen Li

Keyword(s):

Hepatitis A ◽

Hepatitis A Virus ◽

Rt Pcr ◽

Consensus Primer

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Quantification of Hepatitis A Virus in Shellfish by Competitive Reverse Transcription-PCR with Coextraction of Standard RNA

Applied and Environmental Microbiology ◽

10.1128/aem.65.1.322-326.1999 ◽

1999 ◽

Vol 65 (1) ◽

pp. 322-326 ◽

Cited By ~ 19

Author(s):

Charlotte Arnal ◽

Virginie Ferre-Aubineau ◽

Berangere Mignotte ◽

Berthe Marie Imbert-Marcille ◽

Sylviane Billaudel

Keyword(s):

Tissue Culture ◽

Reverse Transcription ◽

Hepatitis A ◽

Hepatitis A Virus ◽

Internal Standard ◽

Wild Type ◽

Rt Pcr ◽

Reverse Transcription Pcr ◽

Reproducible Method ◽

Dna Enzyme Immunoassay

ABSTRACT To quantify hepatitis A virus (HAV) in experimentally contaminated mussels, we developed an internal standard RNA with a 7-nucleotide deletion for competitive reverse transcription (RT)-PCR. Deposited directly into the sample, this standard was used both as extraction control and as quantification tool. After coextraction and competitive RT-PCR, standard and wild-type products were detected by differential hybridization with specific probes and a DNA enzyme immunoassay. The quantifiable range with this reproducible method was 104 to 107 copies of HAV/gram or 400 to 106 50% tissue culture infective doses/ml.

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