Cloning, Expression and Bioinformatic Analysis of Human Papillomavirus Type 52 L1 Capsid Gene from Indonesian Patient

Suhandono, Sony; Ungu, Dewi Ayu Kencana; Kristianti, Tati; Sahiratmadja, Edhyana; Susanto, Herman

doi:10.5454/mi.8.3.2

Cited by 3 publications

(3 citation statements)

References 29 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This is consistent with previous studies conducted in Bandung by Fadhilah with HPV-16 isolates that have origins kinship with isolates from Asia and East Asia (14) and the research conducted by Suhandono on isolate single HPV-52 infection that has real origin kinship with the isolates from the Philippines (Asia). (15) This shows that there are certain types of HPV genomes that were clustered according to the ethnic people and regions where they are isolated. From the results of this study, the isolates in Bandung HPV L1 genes have origins kinship with isolates from EU077219 Canada and KT799980, KU951191.1 Southwest China.…”

Section: Discussionmentioning

confidence: 97%

Phylogenetic Analysis of Human Papillomavirus 16 and 52 L1 Gene from Cervical Cancer in Bandung, Indonesia

et al. 2018

Self Cite

View full text Add to dashboard Cite

BACKGROUND: Chronic infection with high-risk type of human papillomavirus (HPV) can cause cervical cancer. Previous studies showed that multiple infections of HPV are found in cervical cancer caused by multiple HPV infections and the most common are HPV-16 and HPV-52. The origin of HPV-16 circulating in Indonesia varies. The purpose of this study was to explore the origin of multiple infections of HPV-52 and HPV-16 in cervical cancer by using a phylogenetic tree.METHODS: During July-November 2010, 100 women were diagnosed with cervical cancer in the Department of Obstetrics and Gynecology, Dr. Hasan Sadikin General Hospital, Bandung, Indonesia. Only 96 patients were involved in this study. Ninety-six samples of HPV deoxyribonucleic acid (DNA) were isolated from biopsied tissue of cervical cancer. Multiple infections of HPV genotypes HPV-16 and HPV-52 were confirmed by using the linear assay for HPV genotyping test. Afterward,HPV-52L1 gene was amplified by using self-designed primer. L1 gene was also sequenced and analyzed using phylogenetic program (MEGA6.06).RESULTS: The result of phylogenetic tree construction showed that isolated HPV-52 originated from multiple infections of HPV-16 and HPV-52 from cervical cancer patients in Bandung were in a subgroup with isolates originating from EU077219 Canada (America) and KT799980 southwest China (Asia). Isolate HPV-16 in one subgroup with isolates originating from KU951191.1 (Southwest China).CONCLUSION: L1 gene sequence from multiple infections isolated from HPV-16 and HPV-52 from cervical cancer patients in Bandung refers to the variation of L1 gene reported from Canada and southwest China. This proves that Indonesia’s HPV clusters are located in the strains found in America and Asia.KEYWORDS: multiple infections, HPV-16, HPV-52, L1 gene, phylogenetic

show abstract

Section: Discussionmentioning

confidence: 97%

Phylogenetic Analysis of Human Papillomavirus 16 and 52 L1 Gene from Cervical Cancer in Bandung, Indonesia

et al. 2018

Self Cite

View full text Add to dashboard Cite

show abstract

“…8 The L1 gene in the plasmid was the result of cloning from a biopsy sample of a patient with cervical cancer obtained from Dr. Hasan Sadikin Hospital, Bandung. 9 Upon arrival, the plasmid was sequenced with primer MY09 and MY11 to confirm the presence of the HPV52 L1 gene in the plasmid.…”

Section: Methodsmentioning

confidence: 99%

Semi-quantitative Digital Analysis for Human Papillomavirus Detection from Environmental Specimens

Siagian

Pratama

Fathurrohman

et al. 2020

amj

View full text Add to dashboard Cite

Background: Recently, human papillomavirus (HPV) deoxyribonucleic acid (DNA) has been detected in urban wastewater, indicating that the virus can reach the sewer and, eventually, other water environments. This study aimed to develop a semi-quantitative assay for HPV DNA detection from environmental specimens using the PCR gel electrophoresis method. Method: This was an experimental descriptive qualitative study conducted from July to November 2019 in a standard molecular laboratory and non-laboratory administration room without air conditioner. Three brands of PCR reagents and different annealing temperatures were compared to identify the best condition for conventional PCR of plasmid DNA containing the HPV L1 gene. The semi-quantitative data were obtained from densitometry digital analysis using an imaging software. The optimized protocol was then applied on DNA serial dilutions to seek for the lower limit of detection (LLOD) value and the linear range of the assay. To evaluate the robustness of the assay, the protocol was further applied to spiked specimens of wastewater. Finally, several wastewater samples were tested for the presence of HPV DNA using this protocol. Results: A broad linear range and HPV L1 gene detection ability were observed with an LLOD of less than 2pg plasmid DNA in field condition. Although the assay successfully detected HPV DNA from several spiked wastewater samples, certain wastewater could interfere with the assay and gave false negative result. Conclusion: A semi-quantitative conventional PCR method to detect HPVDNA from environmental samples has been established and proven to be robust in field condition with non-optimum cold chain.

show abstract

“…Different plant-based expression systems were considered, from whole plant approaches for transient expression to stably transformed green microalgae, using single HPV antigens or fusion to peptides to improve accumulation yield, to intracellular targeting strategies (Chabeda et al, 2018 (Parkin and Bray, 2006). HPV VLPs have been successfully produced in plants via transient expression to assemble into higher order structures, and elicit anti-L1 and anti-L2 antibody responses that neutralize HPV-16 and HPV-52 pseudovirions (Pineo et al, 2013;Suhandono et al, 2014). Plant virus particles are easily purified, stable at high temperatures, and functionally indistinguishable from VLPs produced in the conventional system, and they have stronger CD8+ T cell immune responses (Lamprecht et al, 2016).…”

Section: Introductionmentioning

confidence: 99%

Expression of Human papillomavirus type 52 L1 capsid gene in Oryza sativa involved in cytoprotective activities

Lin

Pan

Chen

et al. 2020

Not Bot Horti Agrobo

View full text Add to dashboard Cite

Female cervical cancer is largely formed by Human papillomavirus (HPV), the second leading cause of cancer deaths in women worldwide. HPV-52 is a regionally common high-risk type of cervical cancer found mostly in Asia and reveals geographical variations, in order of importance, as types HPV-16 and -18. However, the differing propensities of HPV types in progressing to cancer, focusing on HPV-52 vaccines, are limited. Several plant-based vaccines against cancer have been developed, and the production of candidate HPV therapeutic vaccines using plant-derived expression platforms is also proven. The objectives of this study were to assess the HPV-52L1 Capsid gene by transferring HPV-52L1 Capsid cDNA into rice (Oryza sativa L.) via an Agrobacterium-mediated transformation, and accumulating HPV-52L1 Capsid proteins in a plant-based expression system to maintain and improve antigenicity. Crude protein extracts containing 5~20 μg from OsHP-52L1 transgenic lines induced cell death and significantly reduced cell proliferation in HPV-positive HeLa cervical cancer cells compared with those non-transformant (NT) rice plants. However, no significant cytotoxicity of induced human breast MDA-MB-231 cell proliferation (as negative control) was observed at any dose compared with NT groups. HeLa cells ameliorated the effects of OsHPV crude protein extracts on cell viability as the extract concentration increased, and treatment with 20 μg of the extract from OsHPV-3 significantly reduced cell viability in HeLa cells (26%) compared with the control group (57%). Our results can be used for exploring the potential of plants for increasing the immunogenicity of OsHPV-52L1 Capsid DNA vaccines, and support the development of cost-effective HPV vaccines, which is highly desirable for resource-poor countries.

show abstract

Cloning, Expression and Bioinformatic Analysis of Human Papillomavirus Type 52 L1 Capsid Gene from Indonesian Patient

Cited by 3 publications

References 29 publications

Phylogenetic Analysis of Human Papillomavirus 16 and 52 L1 Gene from Cervical Cancer in Bandung, Indonesia

Phylogenetic Analysis of Human Papillomavirus 16 and 52 L1 Gene from Cervical Cancer in Bandung, Indonesia

Semi-quantitative Digital Analysis for Human Papillomavirus Detection from Environmental Specimens

Expression of Human papillomavirus type 52 L1 capsid gene in Oryza sativa involved in cytoprotective activities

Contact Info

Product

Resources

About