Journal: Vaccines

Article Title: Immunogenicity and Efficacy of a Trivalent HSV-2 gC2, gD2, gE2 Nucleoside-Modified mRNA-LNP Vaccine Against HSV-1 Eye Infection and Neuroinvasion in Mice

doi: 10.3390/vaccines14030253

Figure Lengend Snippet: Inoculation of HSV-1 produces dose-dependent changes in survival, blepharoconjunctivitis, and weight loss. HSV-1 was inoculated onto the intact cornea of each eye. ( A ) Survival curve. p values calculated by the Log-rank test. n = 5 mice/group ( B ) The blepharoconjunctivitis disease score is shown for the most affected eye on the most severe day of disease. n = 5 mice/group. ( C ) Percent weight loss at time of humane euthanasia or at the end of the study on day 14 post-infection. n = 5 mice/group, except n = 4 for the 10 4 group. p values for ( B , C ) were calculated by Ordinary one-way ANOVA and Dunnett’s multiple comparisons test. * p < 0.05, ** p < 0.01, *** p < 0.001; **** p < 0.0001; ns, not significant.

Article Snippet: To determine HSV-1 DNA copy number, we ran purified HSV-1 DNA (ATCC, VR-539DQ) or adipsin DNA (BioChain, Princeton, NJ, USA, D1334149) as standards.

Techniques: Infection

Journal: Vaccines

Article Title: Immunogenicity and Efficacy of a Trivalent HSV-2 gC2, gD2, gE2 Nucleoside-Modified mRNA-LNP Vaccine Against HSV-1 Eye Infection and Neuroinvasion in Mice

doi: 10.3390/vaccines14030253

Figure Lengend Snippet: Kinetics of infection after HSV-1 was placed on the intact cornea of each eye. Animals were infected with 10 5 PFU (1 lethal dose 50, LD 50 ) of HSV-1 in each eye and sacrificed on days 1, 3, 5, and 7 post-infection ( n = 5 per group). Each tissue was processed separately and analyzed for virus titers (blue) and HSV-1 DNA copy number by qPCR (red). The tissues included ( A ) eye (eye bulb and attached ocular conjunctiva), ( B ) trigeminal ganglion, ( C ) brainstem, ( D ) olfactory bulb, and ( E ) cerebrum/cerebellum.

Article Snippet: To determine HSV-1 DNA copy number, we ran purified HSV-1 DNA (ATCC, VR-539DQ) or adipsin DNA (BioChain, Princeton, NJ, USA, D1334149) as standards.

Techniques: Infection, Virus, Olfactory

Journal: Vaccines

Article Title: Immunogenicity and Efficacy of a Trivalent HSV-2 gC2, gD2, gE2 Nucleoside-Modified mRNA-LNP Vaccine Against HSV-1 Eye Infection and Neuroinvasion in Mice

doi: 10.3390/vaccines14030253

Figure Lengend Snippet: Serum IgG ELISA (binding) and neutralizing antibodies. Mice were immunized twice IM with PBS or 10 μg of the trivalent HSV-2 mRNA vaccine. Sera were obtained 4 weeks after the second immunization, before infection with HSV-1. ( A ) Serum IgG ELISA endpoint titers against HSV-2 or cross-reacting antibodies against HSV-1. ( B ) Neutralizing antibodies to HSV-1. n = 9 or 10 animals/group. p values were calculated using the two-tailed Mann–Whitney test. **** p < 0.0001; ns, not significant.

Article Snippet: To determine HSV-1 DNA copy number, we ran purified HSV-1 DNA (ATCC, VR-539DQ) or adipsin DNA (BioChain, Princeton, NJ, USA, D1334149) as standards.

Techniques: Enzyme-linked Immunosorbent Assay, Binding Assay, Infection, Two Tailed Test, MANN-WHITNEY

Journal: Vaccines

Article Title: Immunogenicity and Efficacy of a Trivalent HSV-2 gC2, gD2, gE2 Nucleoside-Modified mRNA-LNP Vaccine Against HSV-1 Eye Infection and Neuroinvasion in Mice

doi: 10.3390/vaccines14030253

Figure Lengend Snippet: Vaccine protection against disease after HSV-1 inoculation on the intact cornea of each eye. Mice were immunized with PBS or 10 μg trivalent mRNA, and 28 days after the second immunization, the intact cornea of each eye was inoculated with 10 6 PFU (10 LD 50 ) HSV-1 ( n = 10/group). ( A ) Survival curve. The p value was determined using the Log-rank test. ( B ) Blepharoconjunctivitis disease score on the most severe day of disease post-infection for each animal. ( C ) Weight loss. p values for ( B , C ) were calculated using the two-tailed Mann–Whitney test. n = 10/group; ** p < 0.01; *** p < 0.001.

Article Snippet: To determine HSV-1 DNA copy number, we ran purified HSV-1 DNA (ATCC, VR-539DQ) or adipsin DNA (BioChain, Princeton, NJ, USA, D1334149) as standards.

Techniques: Infection, Two Tailed Test, MANN-WHITNEY

Journal: Vaccines

Article Title: Immunogenicity and Efficacy of a Trivalent HSV-2 gC2, gD2, gE2 Nucleoside-Modified mRNA-LNP Vaccine Against HSV-1 Eye Infection and Neuroinvasion in Mice

doi: 10.3390/vaccines14030253

Figure Lengend Snippet: Vaccine protection as assessed by virus culture and HSV-1 DNA copy number in tissues after HSV-1 inoculation of the intact cornea of each eye. Mice were immunized twice with PBS or 10 μg of the trivalent HSV-2 mRNA vaccine, and challenged in each eye with 10 6 plaque forming units (PFU) HSV-1 (10 LD 50 ). ( A ) Tissues were harvested 5 days post-infection for viral titers in PBS and vaccine groups ( n = 5/group). ( B ) The same tissues were evaluated for HSV-1 DNA copy number. ( C ) Tissues were harvested 7 weeks post-infection for HSV-1 DNA copy number in mice in the vaccine group ( n = 10). The black symbol indicates the same mouse with HSV-1 DNA in the eye and the trigeminal ganglia. p values were calculated by the two-tailed Mann–Whitney test; * p < 0.05; ** p < 0.01; ns, not significant.

Article Snippet: To determine HSV-1 DNA copy number, we ran purified HSV-1 DNA (ATCC, VR-539DQ) or adipsin DNA (BioChain, Princeton, NJ, USA, D1334149) as standards.

Techniques: Virus, Infection, Two Tailed Test, MANN-WHITNEY

Journal: Vaccines

Article Title: Immunogenicity and Efficacy of a Trivalent HSV-2 gC2, gD2, gE2 Nucleoside-Modified mRNA-LNP Vaccine Against HSV-1 Eye Infection and Neuroinvasion in Mice

doi: 10.3390/vaccines14030253

Figure Lengend Snippet: Representative images of the eyelids and palpebral conjunctivae, and the trigeminal ganglia of mice infected with HSV-1 that were unvaccinated or vaccinated with the trivalent HSV-2 mRNA vaccine. ( a , b ) Evaluation for blepharoconjunctivitis in an immunized animal: The eyelid shows minimal mixed inflammatory cell infiltrates in the conjunctival substantia propria (arrow). No HSV-1 immunolabelling is detected by IHC. H&E ( a ) and IHC for HSV-1 ( b ). ( c , d ) Evaluation for blepharoconjunctivitis in an unimmunized mouse: Blepharoconjunctivitis is visible (arrow) with regional necrosis of the epithelium (asterisk) and strong immunolabeling of the surface and follicular epithelium for HSV-1. H&E ( c ) and IHC for HSV1 ( d ). Scale bar for ( a – d ), 100 µm. ( e , f ) Trigeminal ganglion of an immunized mouse: The trigeminal ganglion shows no significant findings. H&E ( e ) and IHC for HSV-1 ( f ). ( g , h ) Trigeminal ganglion of an unimmunized mouse: Meningitis is present in the meninges surrounding the trigeminal ganglion (arrow), and occasional neurons exhibit strong immunolabeling for HSV-1. H&E ( g ) and IHC for HSV1 ( h ). Scale bar for ( e – h ), 20 µm.

Article Snippet: To determine HSV-1 DNA copy number, we ran purified HSV-1 DNA (ATCC, VR-539DQ) or adipsin DNA (BioChain, Princeton, NJ, USA, D1334149) as standards.

Techniques: Infection, Immunolabeling

Journal:

Article Title: Using a Resequencing Microarray as a Multiple Respiratory Pathogen Detection Assay ▿ †

doi: 10.1128/JCM.01870-06

Figure Lengend Snippet: Analytic sensitivity of microarray-based detection for prototype control strains

Article Snippet: Thus, hybridization to a series of probe sets provides redundant presence/absence information for the organism while also revealing strain-specific single-nucleotide polymorphisms relative to the sequence chosen. table ft1 table-wrap mode="anchored" t5 TABLE 1. caption a7 Organism a Sample type a Strain Sample source b Detection limit (genome copies) c HAdV-4 d DNA RI-67 ATCC 10 2 HAdV-4 vaccine DNA CL68578 NHRC 10 2 HAdV-4FS_Navy DNA NRHC 10 2 HAdV-4FS_AirForce DNA AFIOH 10 2 HAdV-4FS_AirForce Viral particles ADL 10 2 HAdV-5 DNA Adenoid 75 ATCC 10 3 HAdV-7 DNA Gomen ATCC 10 2 HAdV-7 Viral particles Gomen ATCC ND HAdV-7a vaccine DNA 55142 NHRC 10 2 HAdV-7FS_Navy DNA NHRC 10 3 B. anthracis DNA Ames AFIP 10 1 B. anthracis Bacterial cells Sterne CRP ND B. pertussis DNA NHRC 10 2 Chlamydia pneumoniae DNA ABi 10 1 Influenza A virus (H1N1) Viral particles PR/8/34 ABi 10 2 Influenza A virus (H3N2) RNA ADL 10 2 Influenza A virus (H5N1) RNA AFIOH 10 1 Influenza B virus Viral particles B/Lee/40 ABi 10 3 Francisella tularensis DNA SCHU4 ATCC 10 3 F. tularensis Bacterial cells SCHU4 CRP ND Human coronavirus DNA/RNA 229E ATCC 10 3 Human coronavirus Viral particles 229E ATCC ND Human coronavirus DNA/RNA OC43 ATCC 10 3 Human coronavirus Viral particles OC43 ATCC ND Rhinovirus 89 Viral particles 41467 Gallo ATCC 10 3 Lassa virus e Plasmids BlueHeron 10 3 M. pneumoniae DNA AFIP 10 3 M. pneumoniae Bacterial cells NHRC ND N. meningitidis DNA Murray ATCC 10 2 Parainfluenza virus 1 Viral particles C-35 ATCC 10 3 Parainfluenza virus 3 Viral particles C 243 ATCC 10 3 RSV A Viral particles A-2 ATCC 10 3 RSV B Viral particles B WV/14617/85 ATCC 10 2 S. pneumoniae DNA AFIP 10 2 S. pyogenes DNA Rosenbach ATCC 10 3 S. pyogenes Bacterial cells NHRC ND Variola major virus e Plasmids BlueHeron 10 3 Vaccinia virus DNA Lister ABi 10 3 Y. pestis DNA D27 AFIP 10 3 Ebola virus e Plasmids BlueHeron 10 3 Open in a separate window a Samples were generated by mixing purified nucleic acid templates in Tris-EDTA buffer to create 10 6 genome copies/μl of stock solution.

Techniques: Microarray, Control, Virus

Journal:

Article Title: Using a Resequencing Microarray as a Multiple Respiratory Pathogen Detection Assay ▿ †

doi: 10.1128/JCM.01870-06

Figure Lengend Snippet: Differentiation by RPM v.1 of various HAdVs causing febrile respiratory infections

Article Snippet: Thus, hybridization to a series of probe sets provides redundant presence/absence information for the organism while also revealing strain-specific single-nucleotide polymorphisms relative to the sequence chosen. table ft1 table-wrap mode="anchored" t5 TABLE 1. caption a7 Organism a Sample type a Strain Sample source b Detection limit (genome copies) c HAdV-4 d DNA RI-67 ATCC 10 2 HAdV-4 vaccine DNA CL68578 NHRC 10 2 HAdV-4FS_Navy DNA NRHC 10 2 HAdV-4FS_AirForce DNA AFIOH 10 2 HAdV-4FS_AirForce Viral particles ADL 10 2 HAdV-5 DNA Adenoid 75 ATCC 10 3 HAdV-7 DNA Gomen ATCC 10 2 HAdV-7 Viral particles Gomen ATCC ND HAdV-7a vaccine DNA 55142 NHRC 10 2 HAdV-7FS_Navy DNA NHRC 10 3 B. anthracis DNA Ames AFIP 10 1 B. anthracis Bacterial cells Sterne CRP ND B. pertussis DNA NHRC 10 2 Chlamydia pneumoniae DNA ABi 10 1 Influenza A virus (H1N1) Viral particles PR/8/34 ABi 10 2 Influenza A virus (H3N2) RNA ADL 10 2 Influenza A virus (H5N1) RNA AFIOH 10 1 Influenza B virus Viral particles B/Lee/40 ABi 10 3 Francisella tularensis DNA SCHU4 ATCC 10 3 F. tularensis Bacterial cells SCHU4 CRP ND Human coronavirus DNA/RNA 229E ATCC 10 3 Human coronavirus Viral particles 229E ATCC ND Human coronavirus DNA/RNA OC43 ATCC 10 3 Human coronavirus Viral particles OC43 ATCC ND Rhinovirus 89 Viral particles 41467 Gallo ATCC 10 3 Lassa virus e Plasmids BlueHeron 10 3 M. pneumoniae DNA AFIP 10 3 M. pneumoniae Bacterial cells NHRC ND N. meningitidis DNA Murray ATCC 10 2 Parainfluenza virus 1 Viral particles C-35 ATCC 10 3 Parainfluenza virus 3 Viral particles C 243 ATCC 10 3 RSV A Viral particles A-2 ATCC 10 3 RSV B Viral particles B WV/14617/85 ATCC 10 2 S. pneumoniae DNA AFIP 10 2 S. pyogenes DNA Rosenbach ATCC 10 3 S. pyogenes Bacterial cells NHRC ND Variola major virus e Plasmids BlueHeron 10 3 Vaccinia virus DNA Lister ABi 10 3 Y. pestis DNA D27 AFIP 10 3 Ebola virus e Plasmids BlueHeron 10 3 Open in a separate window a Samples were generated by mixing purified nucleic acid templates in Tris-EDTA buffer to create 10 6 genome copies/μl of stock solution.

Techniques:

Journal: Clinical chemistry

Article Title: DMSO increases mutation-scanning detection sensitivity in clinical samples using high resolution melting

doi: 10.1373/clinchem.2015.245357

Figure Lengend Snippet: Work flowchart. The mutation containing genomic DNA is amplified by conventional PCR or full-COLD-PCR, followed by HRM scanning. The enrichment effect before and after full-COLD-PCR is confirmed by ddPCR.

Article Snippet: Human genomic DNA from commercial cell lines SW480 (ATCC no. CCL-228 TM , p.R273H, c.818G>A), HCC1008 (ATCC no. CRL-2320 TM , p.D281H, c.841G>C), PFSK-1 (ATCC no. CRL-2060 TM , p.C275G, c.823T>G) was extracted using the DNeasy TM Blood and Tissue kit (Qiagen, Valencia, CA, USA) following the manufacturer’s protocol.

Techniques: Mutagenesis, Amplification, Co-amplification at Lower Denaturation temperature PCR

Journal: Clinical chemistry

Article Title: DMSO increases mutation-scanning detection sensitivity in clinical samples using high resolution melting

doi: 10.1373/clinchem.2015.245357

Figure Lengend Snippet: Full-COLD-PCR products from cell line DNA for HRM scanning. (A–C) HRM detection without DMSO; (D–F) HRM detection in the presence of 7% DMSO. Three TP53 exon 8 mutations were tested, HCC1008, p.R273H, c.841G>C; SW480, p.R273H, c.818 G>A; PFSK-1, p.C275G, c.823 T>G.

Article Snippet: Human genomic DNA from commercial cell lines SW480 (ATCC no. CCL-228 TM , p.R273H, c.818G>A), HCC1008 (ATCC no. CRL-2320 TM , p.D281H, c.841G>C), PFSK-1 (ATCC no. CRL-2060 TM , p.C275G, c.823T>G) was extracted using the DNeasy TM Blood and Tissue kit (Qiagen, Valencia, CA, USA) following the manufacturer’s protocol.

Techniques: Co-amplification at Lower Denaturation temperature PCR