Polymerase chain reaction detection of Puumala virus RNA in formaldehyde-fixed biopsy material

Heiske, Andreas; Anheier, Bärbel; Pilaski, J.; Klenk, Hans Dieter; Gröne, Hermann Josef; Feldmann, Heinz

doi:10.1046/j.1523-1755.1999.00421.x

Cited by 16 publications

(15 citation statements)

References 27 publications

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“…Recent studies have been able to analyze mRNA from formaldehyde-fixed, paraffinembedded material [Schutze and Lahr, 1998;Fink et al, 2000;Heiske et al, 2000;Kasi et al, 2000;Imamichi et al, 2001;Cohen et al, 2002]. Specht and co-workers reported a protocol for the isolation of mRNA from formaldehyde-fixed, paraffin-embedded tumor tissues via the digestion of protein-RNA crosslinks by extended proteinase K incubation [Schutze and Lahr, 1998;Specht et al, 2001].…”

Section: Discussionmentioning

confidence: 99%

Differential gene expression analysis using paraffin‐embedded tissues after laser microdissection

Kim

Hwang

et al. 2003

J of Cellular Biochemistry

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Recent advances in laser microdissection allow for precise removal of pure cell populations from morphologically preserved tissue sections. However, RNA from paraffin-embedded samples is usually degraded during microdissection. The purpose of this study is to determine the optimal fixative for RNA extractions from laser microdissected paraffin-embedded samples. The integrity of RNA was evaluated with the intactness of 18S and 28S ribosomal RNA by electrophoresis and by the length of individual gene transcripts using RT-PCR. The various fixatives were methacarn (a combination of methanol, chloroform, and acetic acid) and several concentrations of ethanol and isopropanol. Methacarn was the optimal fixative for RNA preservation in paraffin-embedded tissues, which included liver, lung, kidney, muscle, and limb. Based on RT-PCR analysis, methacarn fixed samples exhibited the expected RNA sizes for individual genes such as glyceraldehyde-3-phosphate-dehydrogenase (GAPDH) and bone-related genes (e.g., alkaline phosphatase and osteonectin). The laser microdissection technique with methacarn fixation was then applied to analyze the differential gene expression between hypertrophic and proliferative chondrocytes in the growth plate of long bone. The expression of type X collagen, a specific gene for hypertrophic chondrocytes, was only observed in hypertrophic chondrocytes, while type II collagen was observed more broadly in the growth plate as anticipated. Thus, combining laser microdissection with methacarn fixation facilitates the examination of differentially expressed genes from various tissues.

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Section: Discussionmentioning

confidence: 99%

Differential gene expression analysis using paraffin‐embedded tissues after laser microdissection

Kim

Hwang

et al. 2003

J of Cellular Biochemistry

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“…PCR was performed using protocols established earlier and commonly used in routine diagnostics for hantaviruses (6,14). Briefly, RNA was extracted from heparinized whole blood, blood clots, serum, and urine using different RNeasy kits (Qiagen, Düsseldorf, Germany).…”

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confidence: 99%

“…For urine, 30 ml collected over 24 h was pelleted at ϳ107,000 ϫ g for 2 h (SW41 Beckman rotor) prior to RNA extraction. Amplification of RNA was done by reverse transcription-PCR in a single-step, single-tube reaction using oligonucleotides previously described (6,14).…”

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confidence: 99%

“…In these cases, PCR detection of viral nucleic acid can be performed on blood samples, but a positive result is to be expected only if the samples were taken within the first days after onset of symptoms. Urine and kidney biopsy materials are sometimes better sources for this particular detection assay but are often not available (6,7,14). The first sampling on this case patient was done several days after the appearance of initial clinical symptoms, and therefore, the negative PCR results were not surprising.…”

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confidence: 99%

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Hemorrhagic Fever with Renal Syndrome: Diagnostic Problems with a Known Disease

et al. 2001

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Hemorrhagic fever with renal syndrome (HFRS), caused by different hantaviruses, is a distinct clinical syndrome endemic in several parts of Asia and Europe. However, the clinical picture can sometimes be indistinguishable from that of other infectious or noninfectious diseases. In this report we describe a clinical case, which is a rare occurrence but is a prime example of the difficulties in the diagnosis of HFRS in areas with a low prevalence of the disease

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“…In our previous study, no evidence of PUU virus nucleocapsid protein could be seen in the kidney biopsy specimens of NE patients [12]. PUU virus antigen, however, can be found in blood and urine during the acute phase of NE [14], and recently hantavirus-specific RNA was observed in kidney biopsy specimens of NE patients [15]. Further, it may be noted that the detection or the absence of a given antigen in the glomeruli does not directly show or exclude the possibility that the antigen has a pathogenetic role in glomerular lesions [11].…”

Section: Discussionmentioning

confidence: 99%

Mesangiocapillary Glomerulonephritis Caused by Puumala Hantavirus Infection

Mustonen

Mäkelä

Helin

et al. 2001

Nephron

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Nephropathia epidemica induced by Puumala hantavirus typically causes acute reversible renal function impairment. A typical renal biopsy finding is acute tubulointerstitial nephritis with slight glomerular mesangial changes. We describe here 5 patients who developed the nephrotic syndrome during the convalescent phase of an otherwise typical acute febrile nephropathia epidemica. Renal biopsy of all patients disclosed type I mesangiocapillary glomerulonephritis (MCGN). A clinical remission of the nephrotic syndrome was observed in 4 patients during the follow-up period, and 1 entered into chronic renal failure. Three patients had microscopic hematuria and proteinuria and 2 elevated blood pressure at the latest assessment visit. No patient had clinical or laboratory findings compatible with chronic bacterial, parasitic or viral infections (hepatitis B or C), malignancies, or other disorders known to be associated with MCGN. In conclusion, Puumala hantavirus has to be added to the list of potential agents associated with type I MCGN. Further studies are necessary to establish the incidence of MCGN caused by various hantavirus infections.

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Polymerase chain reaction detection of Puumala virus RNA in formaldehyde-fixed biopsy material

Abstract: The established technique provides a simple and powerful tool that expands the diagnostic possibilities, especially for otherwise unidentified or retrospective cases. It further allows insight into the molecular epidemiology of HFRS-causing agents.

Cited by 16 publications

References 27 publications

Differential gene expression analysis using paraffin‐embedded tissues after laser microdissection

Differential gene expression analysis using paraffin‐embedded tissues after laser microdissection

Hemorrhagic Fever with Renal Syndrome: Diagnostic Problems with a Known Disease

Mesangiocapillary Glomerulonephritis Caused by Puumala Hantavirus Infection

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