ORAL HIV-ASSOCIATED KAPOSI SARCOMA: A COMPARISON BETWEEN IMMUNOHISTOCHEMISTRY AND qPCR TECHNIQUES FOR DETECTION OF HHV8
DOI:
https://doi.org/10.11606/issn.2357-8041.clrd.2015.97224Keywords:
Kaposi Sarcoma, polymerase chain reaction, immunohistochemistryAbstract
Objective: To compare the diagnostic accuracy of immunohistochemistry compared to real-time PCR (using a simple phenol-chloroform DNA extraction protocol) in the detection of HHV8 in small oral biopsies of Kaposi sarcoma. Also to validate the use of this DNA extraction protocol to extract HHV8 DNA.
Material and methods: Seventeen cases of oral KS were examined. Data including gender, age, and anatomic location were obtained from patient´s records and histological sections stained with hematoxylin and eosin (H&E) were reviewed. Immunohistochemistry was used to detect HHV8 in lesions of interest, as well as real-time PCR.
Results: All the patients were HIV positive, the mean age was 35.5 years old, and the affected oral sites were palate (47%), gingiva (29.4%), tongue (11.8%), lip (5.9%), and oral floor (5.9%). Fifteen samples showed positive staining for HHV8 and only two samples were negative. The same results were observed using real-time PCR HHV8-DNA detection.
Relevance: Our findings suggest that immunohistochemistry is faster and cheaper to perform than real-time PCR and was shown to have similar levels of sensitivity and accuracy for detection of HHV8 even in small biopsies. Additionally DNA extraction using a non-commercial kit, as done in this study can further reduce the costs to a pathology service.
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References
Mehta S, Garg A, Gupta LK et al. (2011) Kaposi's sarcoma as a presenting manifestation of HIV. Indian J Sex Transm Dis 32, 108-110.
Khammissa RA, Pantanowitz L, Feller L (2012) Oral HIV-Associated Kaposi Sarcoma: A Clinical Study from the Ga-Rankuwa Area, South Africa. AIDS Res Treat 2012, 873171.
Nakamura S, Salahuddin SZ, Biberfeld P et al. (1988) Kaposi's sarcoma cells: long-term culture with growth factor from retrovirus-infected CD4+ T cells. Science 242, 426-430.
Reis-Filho JS, Souto-Moura C, Lopes JM (2002) Classic Kaposi's sarcoma of the tongue: case report with emphasis on the differential diagnosis. J Oral Maxillofac Surg 60, 951-954.
Chang Y, Cesarman E, Pessin MS et al. (1994) Identification of herpesvirus-like DNA sequences in AIDS-associated Kaposi's sarcoma. Science 266, 1865-1869.
Ablashi DV, Chatlynne LG, Whitman JE, Jr. et al. (2002) Spectrum of Kaposi's sarcoma-associated herpesvirus, or human herpesvirus 8, diseases. Clin Microbiol Rev 15, 439-464.
Mesri EA, Cesarman E, Boshoff C (2010) Kaposi's sarcoma and its associated herpesvirus. Nat Rev Cancer 10, 707-719.
Pauk J, Huang ML, Brodie SJ et al. (2000) Mucosal shedding of human herpesvirus 8 in men. N Engl J Med 343, 1369-1377.
Isola J, DeVries S, Chu L et al. (1994) Analysis of changes in DNA sequence copy number by comparative genomic hybridization in archival paraffin-embedded tumor samples. Am J Pathol 145, 1301-1308.
Lallemand F, Desire N, Rozenbaum W et al. (2000) Quantitative analysis of human herpesvirus 8 viral load using a real-time PCR assay. Journal of clinical microbiology 38, 1404-1408.
Campos MS, Rodini CO, Pinto-Junior DS et al. (2009) GAPD and tubulin are suitable internal controls for qPCR analysis of oral squamous cell carcinoma cell lines. Oral oncology 45, 121-126.
Fatahzadeh M (2012) Kaposi sarcoma: review and medical management update. Oral Surg Oral Med Oral Pathol Oral Radiol 113, 2-16.
Van der Waal IL, J.; Knuutila S. (2005) Kaposi Sarcoma. In Pathology & Genetics Head and Neck Tumors WHO, pp. 193-194. Albany,NY.
Pantanowitz L, Khammissa RA, Lemmer J et al. (2012) Oral HIV-associated Kaposi sarcoma. J Oral Pathol Med.
Coogan MM, Greenspan J, Challacombe SJ (2005) Oral lesions in infection with human immunodeficiency virus. Bull World Health Organ 83, 700-706.
Duus KM, Lentchitsky V, Wagenaar T et al. (2004) Wild-type Kaposi's sarcoma-associated herpesvirus isolated from the oropharynx of immune-competent individuals has tropism for cultured oral epithelial cells. J Virol 78, 4074-4084.
Avon SL, Klieb HB (2012) Oral soft-tissue biopsy: an overview. J Can Dent Assoc 78, c75.
Melrose RJ, Handlers JP, Kerpel S et al. (2007) The use of biopsy in dental practice. The position of the American Academy of Oral and Maxillofacial Pathology. Gen Dent 55, 457-461; quiz 462-453, 488.
Stiller CA, Trama A, Brewster DH et al. (2014) Descriptive epidemiology of Kaposi sarcoma in Europe. Report from the RARECARE project. Cancer Epidemiol 38, 670-678.
Betsem E, Cassar O, Afonso PV et al. (2014) Epidemiology and genetic variability of HHV-8/KSHV in Pygmy and Bantu populations in Cameroon. PLoS Negl Trop Dis 8, e2851.
Chagas CA, Endo LH, Sakano E et al. (2006) Detection of herpesvirus type 8 (HHV8) in children's tonsils and adenoids by immunohistochemistry and in situ hybridization. Int J Pediatr Otorhinolaryngol 70, 65-72.
Pinheiro Rdos S, Ferreira Dde C, Nobrega F et al. (2013) Current status of herpesvirus identification in the oral cavity of HIV-infected children. Rev Soc Bras Med Trop 46, 15-19.
Cattamanchi A, Saracino M, Selke S et al. (2011) Treatment with valacyclovir, famciclovir, or antiretrovirals reduces human herpesvirus-8 replication in HIV-1 seropositive men. J Med Virol 83, 1696-1703.
Rivero ER, Neves AC, Silva-Valenzuela MG et al. (2006) Simple salting-out method for DNA extraction from formalin-fixed, paraffin-embedded tissues. Pathol Res Pract 202, 523-529.
Campos PF, Gilbert TM (2012) DNA extraction from formalin-fixed material. Methods Mol Biol 840, 81-85.
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