Specimen Stability for DNA-based Diagnostic Testing

Farkas, Daniel H.; Drevon, Ann M.; Kiechle, Frederick L.; DiCarlo, Richard G.; Heath, Ellen M.; Crisan, Domnita

doi:10.1097/00019606-199612000-00002

Cited by 26 publications

(9 citation statements)

References 5 publications

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“…In addition to using laboratory resources more efficiently, such batching eliminates run-to-run variations in a PCR which might complicate interpretation of results. Several groups have reported that viral DNA stored for extended periods generally remains positive by qualitative PCR (2,3,8). However, it has not been investigated whether standard storage conditions retain the quantitative information on the viral DNA present in patient specimens.…”

mentioning

confidence: 99%

“…Wiedbrauk and Cunningham (8) showed that HSV DNA can be amplified from cerebrospinal fluid specimens stored at temperatures ranging from 23 to Ϫ72°C over a period of 30 days. Farkas et al (2) used purified DNA stored at 4°C for 3 years to successfully amplify the gene for factor V. However, the same group suggested that frozen swab specimens should be tested within 60 days (3). An important distinction of these earlier studies is that none used real-time detection of the PCR product, relying instead on less-sensitive qualitative detection by ethidium bromide staining of agarose gels.…”

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confidence: 99%

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Quantitative Stability of DNA after Extended Storage of Clinical Specimens as Determined by Real-Time PCR

Jerome

Huang

Wald³

et al. 2002

J Clin Microbiol

134

102

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Viral DNA stored for extended periods can be amplified by PCR. However, it is unknown whether stored specimens give accurate quantitative results by newer real-time PCR techniques. We therefore compared herpes simplex virus DNA levels in specimens before and after 16 months of storage. The levels of viral DNA remained stable whether the DNA was stored as purified DNA or unextracted DNA in a whole specimen.The PCR has proved invaluable in the diagnosis and management of viral diseases by allowing the amplification of minute quantities of viral nucleic acid in patient specimens. PCR has also been instrumental in research on these diseases. A recent advance in PCR technology has been the development of so-called real-time PCR systems. These homogeneous assays allow the rapid detection of the PCR product with a minimum of specimen handling and provide quantitative measurements of the viral nucleic acid in patient specimens.For research purposes, PCR is often performed with specimens which have been stored for an extended period. Even in prospective studies, specimens are often stored for months or years and are then batched for a large PCR run. In addition to using laboratory resources more efficiently, such batching eliminates run-to-run variations in a PCR which might complicate interpretation of results. Several groups have reported that viral DNA stored for extended periods generally remains positive by qualitative PCR (2, 3, 8). However, it has not been investigated whether standard storage conditions retain the quantitative information on the viral DNA present in patient specimens. If storage reduces viral DNA levels as determined by PCR, it would complicate the design of studies which use archived or batched specimens. To address this issue, we performed quantitative PCR for herpes simplex virus (HSV) DNA with a large series of fresh specimens and again after 16 months of storage. Our results demonstrate that the quantitative information for stored specimens is well retained when HSV DNA levels are determined by real-time PCR.

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confidence: 99%

mentioning

confidence: 99%

Quantitative Stability of DNA after Extended Storage of Clinical Specimens as Determined by Real-Time PCR

Jerome

Huang

Wald³

et al. 2002

J Clin Microbiol

134

102

View full text Add to dashboard Cite

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“…DNA extraction and polymerase chain reaction (PCR) amplification were carried out using standard techniques 6 . DNA samples from the families underwent PCR with the fluorescently labelled microsatellite marker D1S2868 (Applied Biosystems PRISM linkage mapping set version 2) and two fluorescently labelled oligonucleotide primers (designed in‐house, and synthesized by Invitrogen TM Life Technologies; available on request) from the region of chromosome 1p21‐22.…”

Section: Methodsmentioning

confidence: 99%

Mutation analysis in Irish families with glomuvenous malformations

O'Hagan

Maloney

Buckley

et al. 2005

British Journal of Dermatology

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Summary Background Glomuvenous malformations (GVMs) are rare bluish lesions that can affect the skin and mucosal surfaces. They represent defects in vasculogenesis. Lesions can occur sporadically or in an autosomal dominant mode of inheritance. Recent studies have shown that mutations in the glomulin gene (GLMN) on chromosome 1p21‐22 are responsible for familial GVMs. Objectives To search for mutations in GLMN in Irish families with GVMs. Methods We identified four Irish families with GVMs and confirmed linkage to chromosome 1p21‐22 in these cases. We sequenced the glomulin gene in all affected and unaffected members of the families. Results Linkage analysis showed that affected individuals from the families shared a common haplotype. Mutation analysis revealed a delAAGAA mutation in exon 3 of the glomulin gene in all four families with GVMs. Conclusions We confirm that mutations in the glomulin gene are responsible for GVMs and suggest a founder Irish mutation in the glomulin gene in four Irish families.

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“…In our experience, once the infrastructure and physician network were built and personnel were trained, then the attention could be focused on the expansion of the collection to other tissue types (eg, preneoplastic lesions and inflammatory tissues) and biological materials (eg, blood, saliva, serum, plasma, and urine). Since the newest molecular techniques require the availability of adequate amounts of high-quality specimens, 15,16 standardized SOPs for the accrual, processing, and storage of tissues had to be established. One of the most important outcomes of the NCI-funded Partnership between PSMHS and MCC is that it allowed the implementation of biobank-related guidelines already proved to be effective at MCC, and the training of staff in tissue biobanking by MCC co-leaders with that expertise.…”

Section: Flores Et Almentioning

confidence: 99%