The duration of fixation influences the yield of HCV cDNA-PCR products from formalin-fixed, paraffin-embedded liver tissue

1994 ◽  
Vol 48 (2-3) ◽  
pp. 267-272 ◽  
Author(s):  
D. Bresters ◽  
M.E.I. Schipper ◽  
H.W. Reesink ◽  
B.D.M. Boeser-Nunnink ◽  
H.T.M. Cuypers
Keyword(s):  
Liver Tissue ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Pcr Products ◽  
Formalin Fixed

scholarly journals Molecular Genotyping of the Human Cystic Echinococcosis in Mazandaran Province, North of Iran

2019 ◽  
Author(s):  
Zeynab HEDAYATI ◽  
Ahmad DARYANI ◽  
Shahabeddin SARVI ◽  
Shirzad GHOLAMI ◽  
Mehdi SHARIF ◽  
...  
Keyword(s):  
Cystic Echinococcosis ◽  
Mazandaran Province ◽  
Northern Iran ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Pcr Products ◽  
North Of Iran ◽  
Control Procedures ◽  
Nad1 Gene ◽  
Formalin Fixed

Background: The larval stage of the tapeworm (cestode) Echinococcus granulosus is the etiological agent of hydatidosis or cystic echinococcosis, which is the zoonotic parasitic disease causing morbidity and mortality in both humans and livestock. Due to a lack of accurate data on the human isolates of E. granulosus in Mazandaran Province, northern Iran, the current study aimed to survey the population genetic pattern of cystic echinococcosis isolated from humans by sequencing the mitochondrial genes of NADH dehydrogenase subunit 1 (nad1). Methods: Overall, 47 formalin fixed paraffin-embedded tissue (FFPT) blocks were collected from patients' files in various pathology departments of Mazandaran Province in Iran from 2003 to 2015. PCR was performed to amplify a 398bp DNA fragment of mitochondrial nad1. PCR products were sequenced by Bioneer Corporation (South Korea), and the resulting data were analyzed via relevant software to determine the genotypes. Results: The nad1 gene was successfully amplified on 10 from all of the E. granulosus isolates. Overall, 66.6% and 33.3% of the isolates in the studied area displayed the G1 and G2-G3 genotypes, respectively. Conclusion: This study may provide the foundation for further studies in revealing the regional transmission patterns and also in designing adequate control procedures.

scholarly journals Limited yield of diagnoses of intrahepatic infectious causes of canine granulomatous hepatitis from archival liver tissue

2012 ◽  
Vol 24 (5) ◽  
pp. 888-894 ◽  
Author(s):  
Rae G. Hutchins ◽  
Edward B. Breitschwerdt ◽  
John M. Cullen ◽  
Sally A. Bissett ◽  
Jody L. Gookin
Keyword(s):  
Infectious Disease ◽  
Liver Tissue ◽  
Disease Diagnosis ◽  
Differential Staining ◽  
Copper Accumulation ◽  
Infectious Agents ◽  
Granulomatous Hepatitis ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Formalin Fixed

Canine granulomatous hepatitis is an uncommon morphologic diagnosis that has been associated with a variety of diseases, including a number of systemic infectious etiologies. Formalin-fixed, paraffin-embedded (FFPE) tissues are typically the only source of liver tissue remaining for additional testing for the presence of infectious disease within granulomas. It is unclear if the more common infectious culprits of granulomatous hepatitis can be identified from such specimens. The aim of the current study was to retrospectively investigate archival FFPE liver tissue from dogs with granulomatous hepatitis for the presence of infectious agents. Semiquantitative analysis of copper accumulation in liver specimens was also performed. Medical records were examined for recorded evidence of systemic infectious disease diagnosis. Formalin-fixed, paraffin-embedded liver was prospectively evaluated for infectious agents via differential staining techniques ( n = 13), eubacterial fluorescent in situ hybridization ( n = 11), and Bartonella polymerase chain reaction assays ( n = 15). An infectious cause of granulomatous hepatitis was not identified within liver tissue from any dog using these diagnostic methodologies. Six out of 25 (24%) dogs were diagnosed with concurrent systemic or localized bacterial infections at the time of presentation. Nine out of 17 (53%) dogs had excessive hepatic copper accumulation when evaluated by a semiquantitative histologic grading scheme or quantitative copper analysis. As definitive infectious causes of granulomatous hepatitis were not identified within archival liver biopsy samples, it was concluded that investigation of infectious etiologies within FFPE liver specimens using these diagnostic approaches may be of low yield.

Detection of Merkel cell polyomavirus in formalin-fixed, paraffin-embedded tissue of Merkel cell carcinoma and its correlation with prognosis

2009 ◽  
Vol 27 (15_suppl) ◽  
pp. 6027-6027
Author(s):  
A. A. Andea ◽  
R. Patel ◽  
S. Ponnazhagan ◽  
S. Kumar ◽  
P. DeVilliers ◽  
...  
Keyword(s):  
Tumor Progression ◽  
Survival Rates ◽  
T Antigen ◽  
Ffpe Tissue ◽  
Vp1 Gene ◽  
Merkel Cell ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Pcr Products ◽  
Formalin Fixed

6027 Background: Polyomaviruses are frequently associated with malignancies in their hosts. Recently, a novel polyoma virus, named Merkel cell polyomavirus (MCV) was identified integrated to the genome of 6 of 10 fresh frozen samples of MCC. These findings suggest an involvement of MCV in the pathogenesis of MCC. Because MCC is a rare tumor, the most readily available tissue is archival formalin-fixed, paraffin-embedded (FFPE) material. In this study we evaluated the presence of MCV in FFPE tissue and correlated its presence with tumor progression. Methods: Representative FFPE specimens from 18 tumors belonging to 14 patients with a diagnosis of MCC were retrieved. Following DNA extraction, we performed PCR using 4 different primer pairs amplifying sequences within the T antigen and VP1 gene of MCV. The identity of the PCR products was confirmed by direct sequencing. For clinical outcome analysis we used our MCC database containing 40 patients from 1997 to 2008 with a median follow-up interval of 20 months (range 1–108 months). Results: We detected MCV amplicons in 8 of 18 analyzed tumors (44.4%) from 7 of 14 cases (50%). PCR products from the T antigen portion of MCV were found in 6 of 18 tumors (33.3%) and from the VP1 gene in 2 of 18 tumors (11.1%). For 3 patients, MCV detection was performed in both the primary and metastatic tumors. In one of these the virus was found in both lesions while in the other 2 only the primary tumor demonstrated the virus. Two- and 5-year survival rates for the MCCs containing the virus were 47.6% and 0% respectively while for the rest of the cases in our database 2- and 5-year survival rates were 61.6% and 40.8% respectively; the difference did not reach however, statistical significance (p = 0.3). Conclusions: We identified MCV sequences in half of the cases analyzed demonstrating the feasibility of this technique in FFPE tissue. We also found that the tumors harboring MCV exhibit a trend for more aggressive behavior compared to the rest of MCCs. These findings suggest that MCV might have an etiologic role in carcinogenesis or tumor progression in MCC. Further studies investigating the biological significance of MCV integration in the human genome and the presence of MCV in other neuroendocrine tumors are warranted. No significant financial relationships to disclose.

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