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Category: Volume 06 Issue 01 January 2018
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Title: Detection and Characterization of Kaposi’s sarcoma Herpes Virus (KSHV) from Archival Tissues fixed in Formalin and Paraffin Wax-Embedded at Kenyatta National Hospital, Kenya

Authors: Rodgers N. Demba, Nathan Shaviya, Matilu Mwau, Walter Mwanda

 DOI:  https://dx.doi.org/10.18535/jmscr/v6i1.157

Abstract

The DNA of Human Herpes virus (HHV-8) has been detected in patients with HIV/AIDS (Human Immunodeficiency Virus/Acquired Immunodeficiency Disease). Kaposi’s sarcoma (KS) is a cancer seen in patients who have developed immunosuppression due to HIV. In order to characterize KS, archived tissues fixed in formalin and paraffin wax-embedded were obtained and three distinct gene region were determined by PCR (Polymerase chain reaction) and sequencing: K1, K15 and ORF75. The results showed that the three targeted gene were distinct and were present in the archived samples. The total number of the archived samples that were screened for the presence of HHV-8 DNA were 81. The study was able to detect K1, K15 and ORF75 gene in all the 81 archived samples. The extraction of DNA was by use of the Gene Read TM DNA FFPE Kit and detected by nested PCR using Qiagen®Taq PCR Core Kit where us the PCR products were isolated using 1% agarose gels. The KS gene products were cloned into pTOP V2-TA vector for the sticky-end and sequenced using M13 primers. The study cloned and sequenced 20 samples for each targeted K1, K15 (P) and ORF75 gene. Good sequences result was obtained 50% (10/20) ORF75, 40% (8/20) K15(P) and 75% (15/20) K1gene.Double gene sequences were also detected. The study can authoritatively state that K1, K15 (P) and ORF75 genes sequences for the KS were detected in the archived samples.

Keywords: Kaposi’s sarcoma,Human Herpes virus (HHV-8), Genotyping, Sequencing

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Corresponding Author

Rodgers N. Demba

School of Health Sciences, Kisii University, P.O Box 408-40200, Kisii, Kenya

University of Nairobi Institute of Tropical and infectious Diseases,

P.O. Box 30197, GPO, Nairobi, Kenya