Detection of human papillomavirus (HPV) L1 gene DNA possibly bound to particulate aluminum adjuvant in the HPV vaccine Gardasil.
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.
Milford Hospital and Milford Molecular Laboratory, 2044 Bridgeport Avenue, Milford, CT 06460, USA. shlee01@snet.net
Sin Hang
SH
Lee
D000277:Adjuvants, Pharmaceutic / Q000737:chemistry*; D000536:Aluminum Hydroxide / Q000737:chemistry; D001483:Base Sequence; D036022:Capsid Proteins / Q000235:genetics*; D017931:DNA Primers / Q000737:chemistry; D004279:DNA, Viral / Q000737:chemistry* / Q000235:genetics; D000068857:Human Papillomavirus Recombinant Vaccine Quadrivalent, Types 6, 11, 16, 18; D052140:Human papillomavirus 11 / Q000235:genetics*; D052161:Human papillomavirus 18 / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D009856:Oncogene Proteins, Viral / Q000235:genetics*; D053918:Papillomavirus Vaccines / Q000737:chemistry* / Q000235:genetics; D010710:Phosphates / Q000737:chemistry; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA
D000277:Adjuvants, Pharmaceutic; D036022:Capsid Proteins; D017931:DNA Primers; D004279:DNA, Viral; C114580:HPV L1 protein, Human papillomavirus; D000068857:Human Papillomavirus Recombinant Vaccine Quadrivalent, Types 6, 11, 16, 18; D009856:Oncogene Proteins, Viral; D053918:Papillomavirus Vaccines; D010710:Phosphates; D000536:Aluminum Hydroxide; C524363:aluminum hydroxyphosphate sulfate