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Detection of EBV in nasopharyngeal carcinoma by quantum dot fluorescent in situ hybridization
Authors:Hong-lei Chen  Jun Peng  Xiao-bo Zhu  Jun Gao  Jing-ling Xue  Ming-wei Wang  He-shun Xia
Affiliation:aDepartment of Pathology, School of Basic Medical Science, Wuhan University, 430071 Wuhan, China;bState Key Laboratory of Virology, Wuhan University, 430072 Wuhan, China;cWuhan Tumor Nanometer Diagnosis Engineering Research Center, 430072 Wuhan, China;dDepartment of Pathology, Hubei Cancer Hospital, 430079 Wuhan, China
Abstract:

Aims

Nasopharyngeal carcinoma (NPC) is a common cancer in Southeast Asia and is frequently associated with Epstein–Barr virus (EBV) infection. The primary aim of this study was to improve the method of EBV detection by exploring quantum dots in FISH detection, and compare QD-based FISH with conventional ISH.

Materials and methods

Biopsy specimens were retrospectively retrieved from 35 NPC patients as paraffin-embedded tissue blocks. QD-FISH was developed to detect the presence of EBV encoded small RNA (EBER) using biotin-labeled EBER oligonucleotide probe indirectly labeled with streptavidin-conjugated quantum dots. Conventional ISH was also performed using a commercial kit to assess concordance between the two methods.

Results

All the 35 NPC cases were nonkeratinizing carcinoma (7 differentiated and 28 undifferentiated subtypes). EBER-positive signals were detected in 91.43% (32/35) and 80% (28/35) cases by QD-FISH and ISH, respectively. There was no significant difference in the number of EBER-positive cases by the two methods. A moderate concordance was found between QD-FISH and ISH for EBER status (κ = 0.55). Four EBER-negative cases by ISH showed EBER-positive signals when detected by QD-FISH.

Conclusions

EBV is closely associated with NPC in Chinese patients. QD-FISH is a novel effective method for EBER detection, and has a moderate concordance with conventional ISH.
Keywords:Abbreviations: NPC, nasopharyngeal carcinoma   EBER, Epstein&ndash  Barr virus encoded small RNA   QD-FISH, quantum dot-based fluorescent in situ hybridization   ISH, in situ hybridization   QDs, quantum dots   FFPE, formalin fixed and paraffin-embedded
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