The polymerase chain reaction and its applications in neuropathology

Love, Seth; Nicoll, James A. R.

doi:10.1111/j.1365-2990.1992.tb00772.x

Cited by 10 publications

(4 citation statements)

References 99 publications

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“…The efforts invested in studying single-band amplification (SBA) by polymerase chain reaction (PCR) have led to defining the components and the reaction conditions, including the annealing temperature, the number and duration of the cycles and the quantity of the template (10,11,22). In some cases, such as screening for several markers in restricted amounts of sample, there is a need to perform a duplex PCR amplification (DA).…”

Section: Introductionmentioning

confidence: 99%

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Quantitative Ratio of Primer Pairs and Annealing Temperature Affecting PCR Products in Duplex Amplification

et al. 1999

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The quantity of PCR products that are simultaneously amplified from two different loci in a duplex amplification (DA) are significantly lower for one of the loci, as compared to identical PCR amplification in separate single-band amplifications (SBA). This difference in amplification probably occurs already after the second cycle of amplification. To further analyze this phenomenon, we tested different reaction conditions, including annealing times, a wide range of temperatures, various quantities of the template, several nucleotide concentrations, different amounts of TaqI DNA Polymerase, number of amplification cycles and various amounts of primers and primers ratio. Changing the ratio between the sets of primers in DA had the most significant effect on the relative levels of amplification of the loci with an optimal ratio of 4:1 in favor of the set of primers used to amplify the underrepresented fragment. The optimal annealing temperatures for the tested sets of primers were identical in SBA and different in DA. Possible reasons for this phenomenon are discussed.

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Section: Introductionmentioning

confidence: 99%

“…Quantitative PCR has the potential to become a useful tool in diagnostics and in research applications that require quantitative results (2,3,8,10,13,22). For example, aneuploidities, in chromosomes 21 and 18, can be detected by the quantitative PCR approach (16,17,22).…”

Section: Introductionmentioning

confidence: 99%

Quantitative Ratio of Primer Pairs and Annealing Temperature Affecting PCR Products in Duplex Amplification

et al. 1999

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“…We conclude that it is possible to survey archival tissue and specifically and reproducibly detect target DNA sequences, even when conventional procedures are used for tissue collection, processing and sectioning. This sensitive method has been applied to human autopsy tissues in rather few studies (18), but has the potential to be more widely useful.…”

Section: Discussionmentioning

confidence: 99%

Detection of Viral DNA in Neonatal Herpes Encephalitis Autopsy Tissues by Solution‐Phase PCR: Comparison with Pathology and Immunohistochemistry

et al. 1993

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To detect DNA sequences of herpes simplex virus (HSV) in neural and non-neural tissue sections in disseminated human neonatal HSV infection, a solution polymerase chain reaction (PCR) protocol was developed which amplified HSV thymidine kinase and host genomic DNA sequences that were hybridized with sequence-specific probes in Southern blots. Serial sections of formalin-fixed, paraffin embedded autopsy tissues were tested by PCR and compared to histology and HSV antigen detection. The sensitivity, specificity and reproducibility of this PCR protocol were determined on uninfected and HSV-infected mouse tissues and on HSV DNA from infected tissue culture cells. Samples estimated to contain as few as 60 copies of preserved HSV DNA target sequence gave a positive PCR result. In nine neonates that died during acute HSV infection, all non-neural tissues and a minority of neural tissues with histological lesions had HSV antigen; when DNA could be amplified, HSV DNA sequences were detected by PCR. Together, these findings indicate a direct role for virus in the pathogenesis of these lesions. In the same cases, some or all brain samples were negative for HSV antigen, but nevertheless had HSV DNA sequences detected by PCR. The possible explanations for this finding are discussed. In one neonate dying seven weeks after birth, HSV sequences were found in brain lesions in the absence of HSV antigen; neither HSV DNA nor antigen were found in non-neural tissues, suggesting a latent HSV infection in brain. It is practical to apply PCR methods to detect minute quantities of viral DNA in formalin-fixed, paraffin embedded autopsy tissues.(ABSTRACT TRUNCATED AT 250 WORDS)

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“…Oligoclonal immunoglobulin G autochthonously produced in CSF was described by Tourtelotte in 1970 [16] and free immunglobulin light chains in CSF were detected by Riberi in 1975 [17]. In 1990, successful PCR with cells obtained from CSF were reported [18]. The first flow cytometry analyses of cells in CSF were performed at the beginning of 1990s [19].…”

Section: Csf Analysismentioning

confidence: 99%

Modern cerebrospinal fluid analyses for the diagnosis of diffuse large B-cell lymphoma of the CNS

Baraniskin

Schroers

2014

CNS Oncol.

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CNS lymphomas represent rare and aggressive variants of extranodal non-Hodgkin's lymphomas, which may present with diverse neurological symptoms and are often diagnostically challenging. Primary CNS lymphomas develop within the CNS and characteristically involve the brain, leptomeninges, eyes and, in rare cases, spinal cord. Secondary CNS lymphomas are characterized by expansion of systemic lymphomas to the CNS. Multimodal investigation of cerebrospinal fluid (CSF) comprises an important component of the diagnostic work-up for patients with suspected CNS lymphomas. Cytopathological examination of the CSF is still regarded as the 'gold standard' for the diagnosis of leptomeningeal malignant disease. However, cytopathology has only a low sensitivity in detecting leptomeningeal lymphoma involvement. Modern technologies including proteochemical and immunophenotypic studies by flow cytometry, and molecular genetic analyses of CSF may increase sensitivity and specificity, therefore, facilitating the diagnosis of CNS lymphomas. This review gives an overview and discussion of the current aspects of CSF analyses in CNS lymphomas.

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The polymerase chain reaction and its applications in neuropathology

Cited by 10 publications

References 99 publications

Quantitative Ratio of Primer Pairs and Annealing Temperature Affecting PCR Products in Duplex Amplification

Quantitative Ratio of Primer Pairs and Annealing Temperature Affecting PCR Products in Duplex Amplification

Detection of Viral DNA in Neonatal Herpes Encephalitis Autopsy Tissues by Solution‐Phase PCR: Comparison with Pathology and Immunohistochemistry

Modern cerebrospinal fluid analyses for the diagnosis of diffuse large B-cell lymphoma of the CNS

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