Dear Shaik Mohammed Irshad

Pls. find the attached files

http://digitalcommons.unl.edu/cgi/viewcontent.cgi?article=1055&context=bioscidiss

http://www.who.int/immunization/hpv/learn/hpv_laboratory_manual__who_ivb_2009_2010.pdf

http://www.jbc.org/content/274/9/5810.abstract

http://www.cytoimmun.de/fileadmin/temp/dateien/download/Melsheimer%20et%20al.pdf

http://www.cdc.gov/vaccines/pubs/pinkbook/hpv.html

https://www.cdc.gov/vaccines/pubs/pinkbook/downloads/hpv.pdf

page 272 in: M. A. Hayat. 2010. Methods of Cancer Diagnosis, Therapy, and Prognosis: General Overviews, Head and Neck Cancer and Thyroid Cancer Springer Science & Business Media,Pp.476. ISBN9048131863, 9789048131860

http://www.cytoimmun.de/fileadmin/temp/dateien/download/Melsheimer%20et%20al.pdf

http://www.cytoimmun.de/fileadmin/temp/dateien/download/Hilfrich%20Hariri%20AQCH%20Final.pdf

http://s3.amazonaws.com/academia.edu.documents/40883661/6151.pdf?AWSAccessKeyId=AKIAJ56TQJRTWSMTNPEA&Expires=1475263620&Signature=IErUE4QZtnTEtyyoEUqltE24THI%3D&response-content-disposition=inline%3B%20filename%3DHuman_papillomavirus_type_11_recombinant.pdf

https://www.wikigenes.org/e/gene/e/1489082.html

http://www.wikigenes.org/e/gene/e/1403388.html

http://jvi.asm.org/content/77/21/11625.short

https://www.ncbi.nlm.nih.gov/gene/1489082

https://www.ncbi.nlm.nih.gov/pubmed/22996373

https://cancerci.biomedcentral.com/articles/10.1186/s12935-015-0206-0

http://online.liebertpub.com/doi/full/10.1089/mab.2015.0084?src=recsys

https://www.researchgate.net/publication/277949084_Observations_on_the_expression_of_human_papillomavirus_major_capsid_protein_in_HeLa_cells

http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2011001200003

http://s3.amazonaws.com/academia.edu.documents/36070124/6480.pdf?AWSAccessKeyId=AKIAJ56TQJRTWSMTNPEA&Expires=1475263141&Signature=Ni6wrrL5g1tMU4FecRmCkku7HUo%3D&response-content-disposition=inline%3B%20filename%3DIdentification_of_Two_Cross-Neutralizing.pdf

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC104976/

https://www.ncbi.nlm.nih.gov/pubmed/9650960

Hoping this will be helpful            

Regards

Prof. Houda Kawas

Article Observations on the expression of human papillomavirus major...

Also

Magnus von Knebel Doeberitz, N. Wentzensen. 2007. Human Papillomaviruses and Cervical Cancer, IOS Press, ISBN1586037684, 9781586037680  Disease markers, ISSN 0278-0240

Regards

Houda

I do not understand why you use or choose PCR method.   I have previously informed from Dr. Makoto Iwaya (former Director of Infectious Diseases Laboratory, National Research Institute for Child Health and Development, Tokyo; my unpublished personal information from him) that PCR method seems to be insatiable method especially to start from scanty DNA (from white blood cells of neonate's).   I have recognized that even the thermo-stable DNA polymerase may also change the Km values during amplification reactions.

The effect of presence of thiol reducing reagents on the changes in Km values of an enzyme (thiol-type enzyme biotinidase) has been already reported by us in "Hayakawa K, Yoshikawa K, Oizumi J, Yamauchi K. Determination of biotinidase activity with biotinyl-6-aminoquinoline as substrate. Methods in Enzymology 279: 435-442, 1997.".

Therefore, I would like to recommend the protein-based quantitative/reliable PDMD method instead (please see two files).

By the way, HPV proteins have not been linked to HCC (hepatocellular carcinoma) at all as assessed by PDMD method.   PDMD method has also indicated that co-infection of HCV and HIV-1 is the origin of HCC (see file HepG2 fucoidan).

03/10/2016

By the way, interestingly HPV is detected in fetal liver cell (Hc; normal); i.e., Minor capsid protein L2 (Human papillomavirus type 56; HPV-56) at highest level of 9.0 μg/mg of tissue protein.   Minor capsid protein L2 is also present in HepG2 (+fucoidan; normal); i.e., Minor capsid protein L2 (Human papillomavirus type 67; HPV-67) at scanty level of 0.09 μg/mg of tissue protein.

Major capsid protein L1 has not been present in our tissue specimens.

HPV is not linked to HCC as follows as assessed by PDMD method.

HepG2 (-fucoidan; cancer state) has Replication protein E1 at 0.90 μg/mg of tissue protein.

HepG2 (+fucoidan; normal) has Regulatory protein E2 and Minor capsid protein L2 at 0.14 and          0.09 μg/mg of tissue protein, respectively.

Pseudo liver cancer (normal) has E7 protein and Probable L2 protein at 1.1 and 08 μg/mg of tissue protein, respectively.

LC tissue (non cancer; with leprosy) has E1 protein, E6 protein, and Probable L1 protein at 0.91, 0.23, and 2.3 μg/mg of tissue protein, respectively.

HCC tissue (cancer; with PBC) has Probable L1 protein at 3.1 μg/mg of tissue protein.

LC tissue (non cancer tissue; survived patient No.6) has E1 protein and E2 protein at 0.35 and 3.7 μg/mg of tissue protein, respectively.

HCC tissue (cancer　tissue; survived patient No.6) has E1 protein and Probable L1 protein at 0.07 and 0.54 μg/mg of tissue protein, respectively.

These results suggest that presence of HPV is not related to HCC.

 04/10/2016

Human papillomavirus (HPV) changes gene expression differently between adults and fetal Hc cells and/or babies aged before 4 months..   

Hc cells (normal fetal liver cells; made from Americans) has Minor capsid protein L2 (HPV-56) at 9.0 μg/mg of tissue protein, and expresses or upregulates Jumonji domain-containing protein 2B (Histone demethylase that demethylates 'Lys-4' and 'Lys-36' of histone H3) at 8.9 μg/mg of tissue protein.

Serum of 4mo Japanese boy (before biotin therapy) has Probable protein E5 (HPV-39) at 5.8 μg/mg of serum protein, and excretes Histone-lysine N-methyltransferase 2B/Trithorax homolog 2 at 5.6 μg/mg of serum protein into his blood.

On the other hand, adult and children (older than 1y) changes gene expression and metabolism of protein kinases; i.e.,

LC tissue (No.6) has E1 and E2 protein (HPV) at 4.1 μg/mg of tissue protein, and expresses Serine/threonine protein kinase receptor R5/ALK-3 at 2.2 μg/mg of tissue protein.

HCC tissue (No.6) has Probable L1 protein (HPV) at 0.48 μg/mg of tissue protein, and expresses Serine/threonine-protein kinase A-Raf/Proto oncogene Pks2 at 0.53 μg/mg of tissue protein.

HCC tissue (with PBC) has Probable L1 protein (HPV) at 2.3 μg/mg of tissue protein, and expresses Tyrosine-protein kinase receptor TEK/HPK-6 at 3 μg/mg of tissue protein.

LC tissue (with leprosy) has E1 and E6 protein (HPV) at 1.23 μg/mg of tissue protein and Probable L1 protein (HPV) at 2.3 (total; 3.5) μg/mg of tissue protein, and expresses Serine/threonine-protein kinase receptor R4/ALK-5/TGF-beta receptor type-1 at 4.4 and G protein-coupled receptor kinase GRK5 at 6.2 μg/mg of tissue protein, respectively.

Normal liver tissue of pseudo-cancer has E7 protein (HPV) at 1.1 μg/mg of tissue protein and Probable L2 protein (HPV) 0.8 μg/mg of tissue protein (Total; 1.9), and expresses Serine/threonine-protein kinase receptor R5 /ALK-3 at 1.7 μg/mg of tissue protein.

HepG2 (15 y, without fucoidan) has Replication protein E1 (HPV) at 0.9 μg/mg of tissue protein, and expresses Serine/threonine-protein kinase Nek5/Never in mitosis A-related kinase 5 at 1.1 μg/mg of tissue protein.

HepG2 (with fucoidan) has Regulatory protein E2 (HPV) at 0.09 μg/mg of tissue protein, and expresses Serine/threonine-protein kinase Krs-1/Serine-threonine-protein kinase 3 at 0.28 μg/mg of tissue protein.

HPV and changed-protein-excretion metabolism into blood; i.e.,

Serum of 1y girl (at 16w of biotin therapy) has Replication protein E1 (HPV-30) at 15.1 μg/mg of serum protein, and excretes Abl interactor 2 at 11.4 μg/mg of serum protein into blood.

Serum of 12mo girl (with common cold) has Replication protein E1 (HPV-30) at 5.3 μg/mg of serum protein, and excretes Testis development protein NYD-SP9/Microfibrillar-associated protein 3-like at 12.4 μg/mg of serum protein into blood.

Serum of 33y male (healthy) has Major capsid protein L1 (This protein is only present in this serum; HPV-15) at 10.5 μg/mg of serum protein, and excretes Microtubule-associated serine/threonine-protein kinase 3 at 28.1 μg/mg of serum protein into blood.

Thank you very much professor for your time and detailed explanation.