PMID- 23078778 OWN - NLM STAT- MEDLINE DCOM- 20130924 LR - 20190109 IS - 1873-3344 (Electronic) IS - 0162-0134 (Linking) VI - 117 DP - 2012 Dec TI - Detection of human papillomavirus (HPV) L1 gene DNA possibly bound to particulate aluminum adjuvant in the HPV vaccine Gardasil. PG - 85-92 LID - S0162-0134(12)00267-X [pii] LID - 10.1016/j.jinorgbio.2012.08.015 [doi] AB - Medical practitioners in nine countries submitted samples of Gardasil (Merck & Co.) to be tested for the presence of human papillomavirus (HPV) DNA because they suspected that residual recombinant HPV DNA left in the vaccine might have been a contributing factor leading to some of the unexplained post-vaccination side effects. A total of 16 packages of Gardasil were received from Australia, Bulgaria, France, India, New Zealand, Poland, Russia, Spain and the United States. A nested polymerase chain reaction (PCR) method using the MY09/MY11 degenerate primers for initial amplification and the GP5/GP6-based nested PCR primers for the second amplification were used to prepare the template for direct automated cycle DNA sequencing of a hypervariable segment of the HPV L1 gene which is used for manufacturing of the HPV L1 capsid protein by a DNA recombinant technology in vaccine production. Detection of HPV DNA and HPV genotyping of all positive samples were finally validated by BLAST (Basic Local Alignment Search Tool) analysis of a 45-60 bases sequence of the computer-generated electropherogram. The results showed that all 16 Gardasil samples, each with a different lot number, contained fragments of HPV-11 DNA, or HPV-18 DNA, or a DNA fragment mixture from both genotypes. The detected HPV DNA was found to be firmly bound to the insoluble, proteinase-resistant fraction, presumably of amorphous aluminum hydroxyphosphate sulfate (AAHS) nanoparticles used as adjuvant. The clinical significance of these residual HPV DNA fragments bound to a particulate mineral-based adjuvant is uncertain after intramuscular injection, and requires further investigation for vaccination safety. CI - Copyright (c) 2012 Elsevier Inc. All rights reserved. FAU - Lee, Sin Hang AU - Lee SH AD - Milford Hospital and Milford Molecular Laboratory, 2044 Bridgeport Avenue, Milford, CT 06460, USA. shlee01@snet.net LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20120830 PL - United States TA - J Inorg Biochem JT - Journal of inorganic biochemistry JID - 7905788 RN - 0 (Adjuvants, Pharmaceutic) RN - 0 (Capsid Proteins) RN - 0 (DNA Primers) RN - 0 (DNA, Viral) RN - 0 (HPV L1 protein, Human papillomavirus) RN - 0 (Human Papillomavirus Recombinant Vaccine Quadrivalent, Types 6, 11, 16, 18) RN - 0 (Oncogene Proteins, Viral) RN - 0 (Papillomavirus Vaccines) RN - 0 (Phosphates) RN - 5QB0T2IUN0 (Aluminum Hydroxide) RN - F41V936QZM (aluminum hydroxyphosphate sulfate) SB - IM MH - Adjuvants, Pharmaceutic/*chemistry MH - Aluminum Hydroxide/chemistry MH - Base Sequence MH - Capsid Proteins/*genetics MH - DNA Primers/chemistry MH - DNA, Viral/*chemistry/genetics MH - Human Papillomavirus Recombinant Vaccine Quadrivalent, Types 6, 11, 16, 18 MH - Human papillomavirus 11/*genetics MH - Human papillomavirus 18/*genetics MH - Humans MH - Molecular Sequence Data MH - Oncogene Proteins, Viral/*genetics MH - Papillomavirus Vaccines/*chemistry/genetics MH - Phosphates/chemistry MH - Polymerase Chain Reaction MH - Sequence Analysis, DNA EDAT- 2012/10/20 06:00 MHDA- 2013/09/26 06:00 CRDT- 2012/10/20 06:00 PHST- 2012/06/15 00:00 [received] PHST- 2012/08/26 00:00 [revised] PHST- 2012/08/26 00:00 [accepted] PHST- 2012/10/20 06:00 [entrez] PHST- 2012/10/20 06:00 [pubmed] PHST- 2013/09/26 06:00 [medline] AID - S0162-0134(12)00267-X [pii] AID - 10.1016/j.jinorgbio.2012.08.015 [doi] PST - ppublish SO - J Inorg Biochem. 2012 Dec;117:85-92. doi: 10.1016/j.jinorgbio.2012.08.015. Epub 2012 Aug 30.