New method for the extraction of viral RNA and DNA from cerebrospinal fluid for use in the polymerase chain reaction assay

Casas, Inmaculada; Powell, Lesley; Klapper, Paul E.; Cleator, Graham M.

doi:10.1016/0166-0934(94)00173-e

Cited by 216 publications

(138 citation statements)

References 30 publications

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“…Cada ejemplar o grupo fue resuspendido en tampón Tris-EDTA, seccionado con bisturí y macerado. La extracción del ADN se realizó por el método del tiocianato de guanidina 18 . El ADN de la sangre completa fue extraído con el AxyPrep Multisource Genomic DNA Miniprep Kit (Axygen Biosciences, EEUU), siguiendo las instrucciones del fabricante.…”

Section: Extracción De áCidos Nucleicosunclassified

Detección molecular de Rickettsia massiliae y Anaplasma platys en garrapatas Rhipicephalus sanguineus y caninos domésticos del municipio de Bahía Blanca (Argentina)

Cicuttin¹,

Vidal

Salvo³

et al. 2014

Rev. chil. infectol.

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Section: Extracción De áCidos Nucleicosunclassified

Detección molecular de Rickettsia massiliae y Anaplasma platys en garrapatas Rhipicephalus sanguineus y caninos domésticos del municipio de Bahía Blanca (Argentina)

Cicuttin¹,

Vidal

Salvo³

et al. 2014

Rev. chil. infectol.

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“…Extraction of viral RNA/DNA The extraction of viral RNA or DNA was followed by a previously described protocol 7 . Briefly, 200 µL of lysis buffer (Guanidine Isothyocianate (GuSCN), Invitrogen (USA) 4 M, 0.5% of N-Lauroylsarcosine salt solution, Fluka (USA), 1 mM dithiotreitol (DTT), Invitrogen (USA), 25 mM sodium citrate, Sigma (USA), 20 µg/tube of glycogen, Sigma (USA), 100 copies of plasmid with pseudo rabies virus (PRV), used as internal control were added to 50 µL of CSF.…”

Section: Methodsmentioning

confidence: 99%

Enterovirus and herpesviridae family as etiologic agents of lymphomonocytary meningitis, Southern Brazil

Vidal

Almeida²,

Messias-Reason³

et al. 2011

Arq. Neuro-Psiquiatr.

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Viral meningitis is a common infectious disease of the central nervous system (CNS) that occurs worldwide. The aim of this study was to identify the etiologic agent of lymphomonocytary meningitis in Curitiba, PR, Brazil. During the period of July 2005 to December 2006, 460 cerebrospinal fluid (CSF) samples with lymphomonocytary meningitis were analyzed by PCR methodologies. Fifty nine (12.8%) samples were positive. Enteroviruses was present in 49 (83%) samples and herpes virus family in 10 (17%), of these 6 (10%) herpes simplex virus, 1 (2%) Epstein Barr virus, 2 (3%) human herpes virus type 6 and 1 (2%) mixed infection of enterovirus and Epstein Barr virus. As conclusion enterovirus was the most frequent virus, with circulation during summer and was observed with higher frequency between 4 to 17 years of age. PCR methodology is an important method for rapid detection of RNA enterovirus and DNA herpesvirus in CSF.

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“…DNA was extracted from 50 L whole blood as described by Casas et al 17 The PCR assay was carried out as described by Cooper et al 18 In brief, the reaction mixture for the PCR comprised 10 mM Tris-hydrochloric acid (pH 8.3), 50 nM potassium chloride, 200 M each deoxyneucleoside triphosphate, 1.5 mM MgCl 2, 0.2 M adenovirus primers ADRJC1 and ADRJC2, 18 1.25 U Taq DNA polymerase (Advanced Biotechnologies, Epsom, United Kingdom), and 5 L appropriate DNA sample or sterile water to make up a final volume of 50 L. The assay was performed on a Programmable Dri-Block Gene Ataq thermal cycler (Pharmacia LKB, Uppsala, Sweden) using one cycle each of 94°C for 7 minutes, 55°C for 1 minute, and 72°C for 1.5 minutes, followed by 40 cycles each of 94°C for 1 minute, 55°C for 1 minute, and 72°C for 1.5 minutes. The amplification products were analyzed by electrophoresis in 6% polyacrylamide gels.…”

Section: Polymerase Chain Reactionmentioning

confidence: 99%

Adenovirus infections following allogeneic stem cell transplantation: incidence and outcome in relation to graft manipulation, immunosuppression, and immune recovery

Chakrabarti

Mautner

Osman

et al. 2002

Blood

396

404

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Adenovirus infections occur in 5% to 21% of patients following stem cell transplantation (SCT), with an associated mortality of up to 50%. However, a lack of prospective studies has hampered further developments in the understanding and management of this infection in the posttransplantation setting. We prospectively studied the incidence and outcome of adenovirus infections after SCT using preemptive screening and a policy of reduction or withdrawal of immunosuppressive therapy if the virus was isolated. The incidence of adenovirus infection was 19.7% (15 of 76), and the virus was isolated exclusively in recipients of T-celldepleted grafts. Patients receiving 50 or 100 mg alemtuzumab in vivo were at the greatest risk of adenovirus infection (45% probability) regardless of donor type, and this was related to the slower lymphocyte recovery. Six (40%) of the 15 adenovirusinfected patients developed adenovirus disease. Severe lymphocytopenia (less than 300/L) at the time of first detection of adenovirus was a major risk factor for development of adenovirus disease (P ‫؍‬ .001). In addition, failure to reduce immunosuppression (P ‫؍‬ .04) and a positive result of adenovirus polymerase chain reaction (PCR) in blood at diagnosis (P ‫؍‬ .01) were both associated with fatal adenovirus disease. On the basis of this study, we recommend active surveillance for adenovirus infection in T-cell-depleted SCT and withdrawal or reduction of immunosuppressive treatment, if possible, in patients with adenovirus infection. Preemptive antiviral therapy is warranted for patients with severe lymphocytopenia or positive blood PCR, and in those in whom immunosuppressive therapy cannot be reduced. (Blood.

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New method for the extraction of viral RNA and DNA from cerebrospinal fluid for use in the polymerase chain reaction assay

Cited by 216 publications

References 30 publications

Detección molecular de Rickettsia massiliae y Anaplasma platys en garrapatas Rhipicephalus sanguineus y caninos domésticos del municipio de Bahía Blanca (Argentina)

Detección molecular de Rickettsia massiliae y Anaplasma platys en garrapatas Rhipicephalus sanguineus y caninos domésticos del municipio de Bahía Blanca (Argentina)

Enterovirus and herpesviridae family as etiologic agents of lymphomonocytary meningitis, Southern Brazil

Adenovirus infections following allogeneic stem cell transplantation: incidence and outcome in relation to graft manipulation, immunosuppression, and immune recovery

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