Storage of bovine viral diarrhoea virus samples on filter paper and detection of viral RNA by a RT-PCR method

Vilček, Štefan; Strojný, Ladislav; Durkovic, B.; Rossmanith, W.; Paton, David

doi:10.1016/s0166-0934(00)00258-5

Cited by 30 publications

(22 citation statements)

References 10 publications

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“…A temperature of Ϫ70°C was optimal for storage. These findings are consistent with those of studies with HIV and hemorrhagic enteritis virus (6,26). The rate of partial degradation of dried DNA on filters or the presence of PCR inhibitors may increase proportionately with storage temperature.…”

Section: Discussionsupporting

confidence: 90%

“…No significant difference in DNA extraction yield was ob- (1,26). Serum dried on filters was stable under different temperatures for 4 weeks, but the yields of PCR products decreased when the temperature was 4°C.…”

Section: Discussionmentioning

confidence: 96%

“…It enables large-scale, worldwide screening of viruses, evolutionary studies, and searches for mutations (22,26). It allows samples from distant field or developing countries, in which cold storage and transportation are problems, to be concentrated into one laboratory (12).…”

Section: Discussionmentioning

confidence: 99%

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Detection of Duck Hepatitis B Virus DNA on Filter Paper by PCR and SYBR Green Dye-Based Quantitative PCR

Wang¹,

Giambrone²,

Smith

2002

J Clin Microbiol

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Duck hepatitis B virus (DHBV) belongs to the Hepadnaviridae family, which includes human Hepatitis B virus (HBV) and Woodchuck hepatitis virus.It is widely distributed in wild and domestic ducks due to congenital transmission. HBV is a worldwide health problem, with carriers at risk of developing cirrhosis and liver cancer. Medical staff and scientists working with HBV must be vaccinated because of its contagious nature. DHBV is a safe surrogate for HBV because of their similarities. Collection of serum and blood samples on filter paper has been used to screen for metabolic disorders, genetic diseases, and viral infection and for evolutionary studies of the genome. In this study, DHBV from serum and blood dried on filters was detected by PCR. A 0.1-l sample was sufficient for detection. The immobilization potential of filter papers for DHBV was examined, and the highest yield of PCR products was observed with Whatman paper. Dried serum was stable under different storage temperatures for 4 weeks, but the yields of PCR products decreased when the temperature was 4°C. The optimal condition for storage was ؊70°C. A newly developed quantitative PCR based on monitoring the amplification by measuring the increase in fluorescence caused by the binding of SYBR green dye to double-stranded products was applied herein. DHBV genomic DNA cloned in a plasmid was used for the generation of standard DHBV DNA for quantitative PCR. It validated results from PCR in terms of the copy number of DHBV particles. The specificity of PCR was demonstrated by melting curve analysis, and the differentiation of two DHBV isolates amplified from dried serum was demonstrated based on their melting temperatures determined by GC contents and sequence. It was easier and simpler than other PCR-based DNA techniques. The use of serum dried on filters allows samples from distant field for which cold storage and transportation are a problem to be mailed to the diagnostic laboratory. Samples can be archived for comparison and used as a source of DNA for cloning and sequencing. Duck hepatitis B virus (DHBV) is the first hepatitis B virus(HBV) identified from an avian host. The natural route of transmission is from the bloodstream of persistently infected laying ducks to the egg, resulting in congenital infection (18,21). Congenitally infected ducks are at risk of developing hepatoma or secondary amyloidosis due to chronic stimulation of the immune system (9, 10, 11). HBV is a worldwide human health problem, with carriers at risk for developing cirrhosis and hepatocarcinoma. The molecular biology of HBV is intensely researched for its medical importance and life cycle. It is dangerous to work with because of nosocomial infections through blood and body fluids from infected patients. Therefore, attention has been devoted to a safe model for HBV such as DHBV (23).Specimens of serum or blood dried on filter paper have been used for serologic and screening tests for human immunodeficiency virus (HIV), measles virus, human cytomegalovirus, phenylketonuria, and...

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Section: Discussionsupporting

confidence: 90%

Section: Discussionmentioning

confidence: 96%

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Detection of Duck Hepatitis B Virus DNA on Filter Paper by PCR and SYBR Green Dye-Based Quantitative PCR

Wang¹,

Giambrone²,

Smith

2002

J Clin Microbiol

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“…Vilcek et al (19) reported no difference in the yield of PCR products of bovine diarrheal virus after storage of dried blood samples at Ϫ20°C, 4°C, and room temperature after 180 days of storage.…”

Section: Resultsmentioning

confidence: 99%

Chromatography Paper Strip Method for Collection, Transportation, and Storage of Rotavirus RNA in Stool Samples

et al. 2004

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We developed a novel method that uses sodium dodecyl sulfate-EDTA-treated chromatography paper strips to collect unconcentrated fresh stool samples. After the paper strips were stored for 4 months at room temperature, rotavirus RNA could be successfully amplified by using reverse transcriptase PCR. The use of filter paper strips as a specimen support allows (self-)collection of stool samples by untrained persons. Diarrheal stool samples from remote areas can be stored and transported to a central diagnostic laboratory without the need for freezers or special shipping conditions. This convenient and inexpensive rotavirus sample collection system can be of use in epidemiological surveillance studies and vaccine trials.

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“…(1,5,11,17,23,24,25,26,30,36). The stability and molecular analysis of DNA in specimens stored on FTA Ò have been extensively documented (38), but little is known about the stability of viral RNA.…”

mentioning

confidence: 99%

Molecular Analysis of Infectious Bursal Disease Virus from Bursal Tissues Collected on FTA® Filter Paper

2006

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We investigated the feasibility of using FTA filter cards for the storage of bursas of Fabricius containing infectious bursal disease virus (IBDV) and for IBDV detection by reverse transcriptase (RT)-polymerase chain reaction (PCR), and characterization by restriction fragment length polymorphism (RFLP) or nucleotide sequencing. The FTA card is a cotton-based cellulose membrane containing lyophilized chemicals that lyses many types of bacteria and viruses. IBDV was inactivated upon contact with the FTA as shown by the inability of the virus to be propagated in embryonating chicken eggs. Viral RNA in minced bursas or stamped bursas could be amplified by RT-PCR (VP2 gene fragment, 248 base pairs) after storage on FTA for at least 15 days at room temperature or 8 mo at -20 C. Analytical sensitivity of the test was between 0.5-5 ng of RNA template or 5 x 10(1) mean tissue culture infective dose (TCID50)/FTA spot. Detection rate of IBDV in domestic clinical samples collected on FTA or collected by the non-FTA standard procedure was 36.7% and 41.7%, respectively, which represents 88% agreement. Detection of IBDV from FTA cards inoculated with bursal tissues in the laboratory or in the field was 36.7% and 37.1%, respectively. Detection of IBDV from FTA samples when the cards were inoculated with bursal tissues and sent through customs into the United States was 32.9%. Analysis of the amplified products showed that molecular characterization of IBDV by RFLP or nucleotide sequencing is feasible in bursas stored on FTA at 25 C for 1-3 mo or at -20 C for at least 8 mo. The use of FTA for the collection of bursal tissues and simultaneous inactivation of IBDV allows the movement of specimens within the United States and also from outside the United States in compliance with federal regulations and in a manner adequate for molecular characterization.

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Storage of bovine viral diarrhoea virus samples on filter paper and detection of viral RNA by a RT-PCR method

Cited by 30 publications

References 10 publications

Detection of Duck Hepatitis B Virus DNA on Filter Paper by PCR and SYBR Green Dye-Based Quantitative PCR

Detection of Duck Hepatitis B Virus DNA on Filter Paper by PCR and SYBR Green Dye-Based Quantitative PCR

Chromatography Paper Strip Method for Collection, Transportation, and Storage of Rotavirus RNA in Stool Samples

Molecular Analysis of Infectious Bursal Disease Virus from Bursal Tissues Collected on FTA® Filter Paper

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