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

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.

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Genotype characterization of livestock and human cystic echinococcosis in Mazandaran province, Iran

Journal of Helminthology ◽

10.1017/s0022149x1800010x ◽

2018 ◽

Vol 93 (2) ◽

pp. 255-259 ◽

Cited By ~ 5

Author(s):

T. Gorgani-Firouzjaee ◽

N. Kalantrai ◽

S. Ghaffari ◽

J. Alipour ◽

S. Siadati

Keyword(s):

Cystic Echinococcosis ◽

Genotypic Diversity ◽

Cox1 Gene ◽

Mazandaran Province ◽

Major Step ◽

Northern Iran ◽

Formalin Fixed Paraffin ◽

Pcr Rflp ◽

Control Programmes ◽

Formalin Fixed Paraffin Embedded

AbstractEchinococcus granulosus is a helminth from the family Taeniidae, which causes cystic echinococcosis (CE) in humans and diverse livestock around the world. The identification of existing genotypes in different regions is a major step towards the prevention and establishment of control programmes for the disease. This study aimed to detect CE genotypes using polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) of the internal transcribed spacer-1 (ITS1) gene and sequencing of the cytochrome c oxidase subunit 1 (Cox1) gene in isolates from the central part of Mazandaran province, northern Iran. Forty isolates were collected from sheep, 17 from cattle and 6 from human formalin-fixed paraffin-embedded tissues (FFPE). The ITS1 and Cox1 genes were successfully amplified by PCR in 41 and 42 samples, respectively. PCR-RFLP and sequencing showed that all isolates had the G1–G3 genotypes in this study. Out of 31 isolates subjected to sequencing for the Cox1 gene, 80.7% had the G1 genotype. G2 (16.1%) and G3 (3.2%) genotypes were observed in five sheep and one cattle samples, respectively. Five human isolates were also sequenced for the ITS1 gene, which showed that all samples belonged to the G1 genotype. Ten haplotypes were determined among the isolates by alignment analysis of the Cox1 gene. In summary, this study demonstrated that G1 was the dominant genotype circulating between humans and livestock in the studied region. Furthermore, high genotypic diversity among the CE isolates was observed.

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

Journal of Virological Methods ◽

10.1016/0166-0934(94)90125-2 ◽

1994 ◽

Vol 48 (2-3) ◽

pp. 267-272 ◽

Cited By ~ 57

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

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

Journal of Clinical Oncology ◽

10.1200/jco.2009.27.15_suppl.6027 ◽

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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