Journal: PLoS Pathogens

Article Title: Hidden regulation of herpes simplex virus 1 pre-mRNA splicing and polyadenylation by virally encoded immediate early gene ICP27

doi: 10.1371/journal.ppat.1007884

Figure Lengend Snippet: ( A ) A diagram of LAT gene structure relative to LAT introns is shown at the top. A a novel splice junction mapped to LAT , flanked by a 1.68 kb “GC-AG” intron is labeled in red. The primary LAT is transcribed antisense to ICP0 , ICP34 . 5 and ICP4 . Relative locations of primers and probes used are labeled under the diagram. The novel splice junction flanked by the 1.68 kb intron and LAT splice junction flanked by the 1.96 kb intron were confirmed by RT-PCR and subsequent sequencing using the cDNAs prepared from HEK-293 cells infected with KOS cells (7 hpi) (Middle of the panel). A previously reported splice junction within the LAT intron (119628^120192) was not confirmed by RT-PCR in the same cDNA sample. Northern blot using two different probes in Vero cells infected with KOS or d27-1 (16 hpi) detected the 1.96 kb LAT intron; however, the 1.45 kb intron and the predicted 1.68 kb intron were under the detection limit. ( B ) Relative splicing efficiency of the three LAT splicing variants were quantitively analyzed by mapping the RNA-Seq data obtained from infected HEK-293 cells to reference sequences including exon-exon and exon-intron junction sequences. No reads mapping to LAT exon-exon and exon-intron junctions were identified for d27-1 infected HEK-293 cells at 4 hpi. Reads mapping to the previously reported splice junction 119628^120192 were not identified in any samples. ( C ) Confirmation of alternative splice junctions for ICP0 . Diagram of novel splice sites (labeled in red) mapping to ICP0 intron 1 and intron 2 (top of the panel). The novel splice variants for ICP0 intron 1 and intron 2 were confirmed by RT-PCR and subsequent sequencing using the same cDNAs prepared from HEK-293 cells infected with KOS cells (7 hpi) (Bottom of the panel). For ICP0 intron 2, the ratio of splice isoforms 122520^122380 (based on the NCBI HSV-1 reference sequence JQ673480) vs. 122520^122377 was approximately 1: 3 in a total of 12 clones sequenced from the PCR band obtained in d27-1 infected cells. Quantitative analysis of usage of the novel 3’ss of ICP0 intron 2 using the RNA-Seq data obtained from infected HEK-293 cells and CLC Genomic Workbench is shown in Panel ( D ). Usage of the novel ICP0 3’ss appear not to be affected by the presence or absence of ICP27. ( E ) Diagram showing novel splice junctions in the UL15 region. Novel splice sites are labeled in red. UL16 and UL17 are transcribed antisense to the UL15 intron (29990^33581). Arrows indicate the relative location of primers used in Panel (G). Quantitative analysis of splicing efficiency of the splice junctions mapping to the UL15 region using the HEK-293 RNA-Seq is shown in Panel ( F ). Presence of ICP27 significantly reduces the splicing efficiency at these novel splice sites. ( G ) Confirmation of novel alternative splice sites mapping to the UL15 region using RT-PCR in HEK-293 cells infected with HSV-1 KOS strain (WT) or d27-1 using primer sets illustrated in panel (E). RT-PCR bands representing novel splicing sites were further confirmed by topo-cloning and DNA sequencing.

Article Snippet: Anti-HSV ICP4 antibody (Santa Cruz) and anti-Flag antibody (Sigma) were sourced commercially.

Techniques: Labeling, Reverse Transcription Polymerase Chain Reaction, Sequencing, Infection, Northern Blot, RNA Sequencing, Clone Assay, Cloning, DNA Sequencing

Journal: PLoS Pathogens

Article Title: Hidden regulation of herpes simplex virus 1 pre-mRNA splicing and polyadenylation by virally encoded immediate early gene ICP27

doi: 10.1371/journal.ppat.1007884

Figure Lengend Snippet: ( A ) Diagram of the ICP34.5 and ICP0 gene locus. Potential ICP34 . 5 mRNA isoforms I-XV, based on identified splice sites, with primers and probe used to detect them. “*” represents pre-mature stop codons in frame with the ICP34 . 5 start codon. Relative locations of two latently expressed viral miRNAs, miR-H3 and miR-H4 , also named miR-I homolog and miR-LAT-ICP34.5 , are labeled on top of the diagram. Novel splice site positions are numbered in red and major novel splice sites (determined by the results presented below) are shown in bold red. ( B ) Confirmation of novel splice junctions mapping to the ICP34 . 5-ICP0 region. cDNAs were prepared from total RNAs from HEK-293 cells infected with KOS or d27-1 at 7 hpi. The same cDNA samples were amplified with different RT-PCR primer sets illustrated in Panel (A). All the novel splice junctions in this region were confirmed by sequencing except for 125650^123186 and 125650^124073, likely due to relatively low levels and/or high GC content in the region. ( C ) Expression of HSV-1 ICP34.5 protein requires ICP27 . Total proteins were prepared from HEK-293 cells infected with KOS or d27-1 at 7 hpi. ICP34.5 was detected using an anti-HSV-1 ICP34.5 antibody, and the same membrane was incubated with an anti-HSV-1 ICP4 antibody, after stripping. ( D ) Splicing using the novel splice sites mapping to the ICP34 . 5-ICP0 region is only efficient when ICP27 is absent. Relative splicing efficiency for the 5 novel splice junctions as well as the ICP0 intron 1 splice variant were quantitively analyzed using the high throughput sequencing data obtained from infected HEK-293 cells. 125650^124046 is the most efficient ICP34.5 splice junction in d27-1 infected cells. ( E ) Detection of ICP34 . 5 splice isoforms in infected HEK-293 cells by Northern blot, with expected sizes of isoforms predicted in Panel (A). Total RNAs were prepared from HEK-293 cells infected with KOS and ICP27 mutant viruses illustrated in the left panel. The probe mapped to exon 1 of ICP34 . 5 is illustrated in Panel A. The 28S and 18S RNA shown in the bottom of the panel were used as loading control. The most abundant isoforms based on Panel D and are labeled in red and confirmed by the blot. ( F ) Detection of ICP34 . 5 isoforms in infected Vero cells by Northern blot, with expected sizes of isoforms predicted in Panel (A). Total RNAs were prepared from Vero cells infected with KOS and d27-1 at 8 hpi or 18 hpi. The 28S and 18S RNA shown in the bottom of the panel were used as loading control. ( G ) Quantitative splicing efficiency analysis for the 5 novel splice junctions mapping in the ICP34.5 region and ICP0 intron 1 splice variants in infected Vero cells. The mean relative splicing efficiency and standard deviation were calculated using the high throughput sequencing data obtained from infected Vero cells.

Article Snippet: Anti-HSV ICP4 antibody (Santa Cruz) and anti-Flag antibody (Sigma) were sourced commercially.

Techniques: Labeling, Infection, Amplification, Reverse Transcription Polymerase Chain Reaction, Sequencing, Expressing, Membrane, Incubation, Stripping Membranes, Variant Assay, Next-Generation Sequencing, Northern Blot, Mutagenesis, Control, Standard Deviation

Journal: PLoS Pathogens

Article Title: Hidden regulation of herpes simplex virus 1 pre-mRNA splicing and polyadenylation by virally encoded immediate early gene ICP27

doi: 10.1371/journal.ppat.1007884

Figure Lengend Snippet: RNA sequences from HEK-293 cells infected with an HSV-1 ICP27 deletion mutant (d27-1) or its wild-type parental strain (KOS) in the presence of the viral polymerase inhibitor phosphonoacetic acid (PAA) or not at 4 and 7 hpi were aligned to the HSV-1 genome (after removal of terminal repeat sequences, which are represented by internal repeats) and graphed as number of viral reads at each genome location. Genome positions of HSV genes relative to the trimmed genome are shown under the graph. Expression of HSV-1 IE genes including RL2 ( ICP0 ), RS1 ( ICP4 ), US1 ( ICP22 ) and US12 ( ICP47 ) labelled in red was similar between KOS or d27-1 infected cells. IE gene UL54 (ICP27) is not detectable in d27-1 infected cells since the coding region of UL54 was deleted in d27-1.

Article Snippet: Anti-HSV ICP4 antibody (Santa Cruz) and anti-Flag antibody (Sigma) were sourced commercially.

Techniques: Infection, Mutagenesis, Expressing

Journal: bioRxiv

Article Title: In HSV-1, the LAT Enhancer Drives Pre-IE VP16 Transcription to Initiate Reactivation

doi: 10.64898/2026.01.06.697999

Figure Lengend Snippet: 17ΔA gene expression following the application of reactivation stimulus wortmannin. A. wt and 17ΔA HSV-1 genome copies in quiescently infected LUHMES cells (MOI of 0.1). Relative genome load was determined by qPCR copy number of DNA polymerase normalized to that of host GAPDH. (n=3). Statistical significance between wt and 17ΔA was calculated by unpaired two-tailed Student’s t-tests with equal variance. B. VP16 gene expression in 17ΔA was quantified by qRT-PCR in LUHMES cells. Cells were quiescently infected at an MOI of 0.2 and total RNA was extracted at 8 dpi (latency), 1, 3, 6 and 24 hpr. Gene expression was quantified using primers and probes listed in Table S2. Relative values for each gene were normalized to host GAPDH expression and were plotted as fold change in expression relative to latency (set to 1). n=3. * p <0.05, ** p <0.005, *** p <0.0005 following Student’s t-tests. C . Expression of representative genes from each kinetic class were quantified by RT-qPCR as described in panel B. D . Comparison between gene expression of wt and 17ΔA at 1 and 6 hpr. E. Luciferase reporter constructs were generated to test the activation of VP16 promoter in neurons using the promoterless pGL3 basic vector, pGL3 basic-LTE (n.t.118,889-119,478), VP16 promoter only, VP5 promoter only, and VP16 or VP5 promoter inserted into the LTE plasmid. F. Luciferase reporter assays were performed in mice N2a cells. All transfections were completed in triplicate wells and were repeated three times biologically. All luciferase values were normalized to the pGL3 basic vector (set to 1). Statistics were determined by unpaired two-tailed Student’s t-tests with unequal variance.

Article Snippet: Viral genome copy numbers were determined by qPCR using primers and probe specific for HSV-1 DNA polymerase (Table S2). qPCRs were performed using TaqManTM Fast Universal PCR Master Mix (2×), no AmpEraseTM UNG (Applied biosystems 4352042) on an Agilent AriaMx Real-Time PCR machine.

Techniques: Gene Expression, Infection, Two Tailed Test, Quantitative RT-PCR, Expressing, Comparison, Luciferase, Construct, Generated, Activation Assay, Plasmid Preparation, Transfection

Journal: bioRxiv

Article Title: In HSV-1, the LAT Enhancer Drives Pre-IE VP16 Transcription to Initiate Reactivation

doi: 10.64898/2026.01.06.697999

Figure Lengend Snippet: Luciferase reporter constructs were generated to test the enhancer-blocking ability of the predicted CTCF insulator sites. A . Schematic representation of CTCFBSDB predicted CTCF binding sites in the UL region of the HSV-1 genome near VP16. B . Representative orientations for each of the constructs tested include the commercially available pGL3-control vector with an SV40 promoter and a luciferase gene as the plasmid backbone; the LTE inserted into the control vector; each putative CTCF-binding site identified in Table S1 together with ∼150-350 bp flanking sequences inserted downstream of the LTE. C . Luciferase reporter assays were performed in N2a as a representative neuronal cell line. All transfections were completed in triplicate. Data represent at least three biological replicates. All luciferase values were normalized to the pGL3-control vector as a fold change in expression relative to control (set to 1). CTRL2 served as a positive control with known LTE-blocking activity in N2a. Statistical comparisons were done on fold changes between the LTE construct and the LTE-CTCF site. * p <0.05, ** p <0.005, *** p <0.0005 by unpaired two-tailed Student s t-tests with unequal variance.

Article Snippet: Viral genome copy numbers were determined by qPCR using primers and probe specific for HSV-1 DNA polymerase (Table S2). qPCRs were performed using TaqManTM Fast Universal PCR Master Mix (2×), no AmpEraseTM UNG (Applied biosystems 4352042) on an Agilent AriaMx Real-Time PCR machine.

Techniques: Luciferase, Construct, Generated, Blocking Assay, Binding Assay, Control, Plasmid Preparation, Transfection, Expressing, Positive Control, Activity Assay, Two Tailed Test

Journal: bioRxiv

Article Title: In HSV-1, the LAT Enhancer Drives Pre-IE VP16 Transcription to Initiate Reactivation

doi: 10.64898/2026.01.06.697999

Figure Lengend Snippet: CTCF and cohesin proteins are enriched on CTUL1 during HSV-1 latency but evicted by 2 hpr. Protein binding was determined by ChIP or CUT&RUN and quantified by qPCR using primers and probes specific to the given gene regions of HSV-1 (Table S2). All relative values for CTCF enrichment were normalized to those of the non-specific binding control IgG and were plotted as fold enrichment relative to IgG (set to 1). n=3-5. A. LUHMES cells were infected at MOI of 0.3 and harvested at 8 dpi for ChIP. CTCF binding in LUHMES was validated at the host positive control region H19/Igf2. qPCR primers specific to CTUL1 or CTRL2 were used to quantitate CTCF enrichment (Table S2). gC served as a negative control of CTCF binding on HSV-1 genome. Each ChIP assay represented one 6-well plate of cells. n=3. Binding significance was determined by Student’s t-tests. * p <0.05. B. CUT&RUN-qPCR was performed with quiescently infected LUHMES cells as described above. CTCF binding significance was determined by one-way analysis of variance (ANOVA). n=3. C. ChIP-qPCR was done with the anti-STAG2 antibody on latently infected LUHMES. STAG2 binding in LUHMES was validated at a host positive control region on human Chromosome 1, as previously reported (Table S2). Primers specific for CTUL1 were used to quantitate STAG2 on each site following ChIP. n=4. gC served as a negative control on the viral genome without STAG2 enrichment. Binding significance was determined by unpaired two-tailed Student’s t-tests. * p <0.05, ** p <0.005. D . LUHMES cells were infected at MOI of 0.3 for 8 days and subjected to wortmannin treatment for 2 hours, followed by ChIP-qPCR. CTCF bound/input values at CTUL1 and CTRL2 were normalized to bound/input at host H19/Igf2. Then, fold change was calculated between 2 hpr and latency (set to 1). * p <0.05, ** p <0.005 by Student’s t-test. E . ChIP-qPCR done with anti-STAG2 antibody on LUHMES subjected to wortmannin treatment for 2 hours. Fold change values plotted were calculated by setting latency to 1. n=3. F . Luciferase reporter constructs were generated to test CTUL1 as an enhancer-blocker to the VP16 promoter in the presence of the CTUL1 insulator binding site and the LTE. A 294-bp random sequence was inserted in place of CTUL1. G. Transient transfections were performed in N2a cells. All transfections were completed in triplicate wells and were repeated three times biologically. All luciferase values were normalized to the pGL3-basic vector (set to 1). * p< 0.05 by unpaired two-tailed Student’s t-tests with unequal variance.

Article Snippet: Viral genome copy numbers were determined by qPCR using primers and probe specific for HSV-1 DNA polymerase (Table S2). qPCRs were performed using TaqManTM Fast Universal PCR Master Mix (2×), no AmpEraseTM UNG (Applied biosystems 4352042) on an Agilent AriaMx Real-Time PCR machine.

Techniques: Protein Binding, Binding Assay, Control, Infection, Positive Control, Negative Control, ChIP-qPCR, Two Tailed Test, Luciferase, Construct, Generated, Sequencing, Transfection, Plasmid Preparation

Journal: bioRxiv

Article Title: In HSV-1, the LAT Enhancer Drives Pre-IE VP16 Transcription to Initiate Reactivation

doi: 10.64898/2026.01.06.697999

Figure Lengend Snippet: A model illustrating how CTCF insulators regulate VP16 de novo gene expression upon HSV-1 reactivation. Beige line indicates HSV-1 genome. Rectangles on the genome denote elements regulatory elements: blue: LAT enhancer; red/green: VP16 promoter; yellow: insulators. Red line denotes transcriptional repression while green line denotes activation. In latency, CTUL1 and another unidentified insulator are enriched with CTCF protein and anchored by the cohesin complex. This structure insulates LTE from turning on the VP16 promoter. In reactivation, CTCF proteins evict from the insulators and cohesin moves away. The loss of enhancer-blocking activity allows LTE to turn on the VP16 promoter.

Article Snippet: Viral genome copy numbers were determined by qPCR using primers and probe specific for HSV-1 DNA polymerase (Table S2). qPCRs were performed using TaqManTM Fast Universal PCR Master Mix (2×), no AmpEraseTM UNG (Applied biosystems 4352042) on an Agilent AriaMx Real-Time PCR machine.

Techniques: Gene Expression, Activation Assay, Blocking Assay, Activity Assay

Journal: Clinical and Experimental Immunology

Article Title: Enhanced expression of IFI16 and RIG‐I in human third‐trimester placentas following HSV‐1 infection

doi: 10.1111/cei.13143

Figure Lengend Snippet: Comparison of the levels of herpes simplex type 1 (HSV‐1) replication in third‐trimester human placentas. Villous and decidual explants were infected with laboratory strain of HSV‐1 in vitro. HSV‐1 DNA was detected and quantified in the placental tissues using the HSV‐1 polymerase chain reaction (PCR) kit using quantitative real‐time (qRT)–PCR under the following conditions: denaturation at 95°C for 10 min, followed by 45 cycles of 95°C for 5 s and 60°C for 40 s.

Article Snippet: Assessment of HSV‐1 load HSV‐1 DNA was detected and quantified in the placental tissues using the HSV‐1 polymerase chain reaction (PCR) kit (GeneProof a.s., Brno, Czech Republic), according to the manufacturer's instructions.

Techniques: Comparison, Infection, In Vitro, Polymerase Chain Reaction, Quantitative RT-PCR

Journal: Clinical and Experimental Immunology

Article Title: Enhanced expression of IFI16 and RIG‐I in human third‐trimester placentas following HSV‐1 infection

doi: 10.1111/cei.13143

Figure Lengend Snippet: The mRNA expression levels of (a) IFI16, (b) TLR3, (c) DDX58 and (d) IFIH1 in third‐trimester mock‐infected and herpes simplex type 1 (HSV‐1)‐infected decidual and chorionic villi tissues (n = 8). Quantitative real‐time polymerase chain reaction (qRT–PCR) was performed using YWHAZ as a control gene. The data are reported as the mean values ± standard error of the mean (s.e.m.). Statistically significant differences compared to mock‐infected placental explants are shown. The Mann–Whitney two‐tailed test was used to assess statistically significant differences.

Article Snippet: Assessment of HSV‐1 load HSV‐1 DNA was detected and quantified in the placental tissues using the HSV‐1 polymerase chain reaction (PCR) kit (GeneProof a.s., Brno, Czech Republic), according to the manufacturer's instructions.

Techniques: Expressing, Infection, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, MANN-WHITNEY, Two Tailed Test

Journal: Clinical and Experimental Immunology

Article Title: Enhanced expression of IFI16 and RIG‐I in human third‐trimester placentas following HSV‐1 infection

doi: 10.1111/cei.13143

Figure Lengend Snippet: Differential expression of (a) IFI16, (b) Toll‐like receptor (TLR)‐3, (c) RIG‐I‐like receptor (RIG‐I) and (d) melanoma differentiation‐associated protein 5 (MDA5) in the placental tissue samples from mock‐infected and herpes simplex type 1 (HSV‐1)‐infected decidua and chorionic villi (n = 8). Data are reported as the mean values ± standard error of the mean (s.e.m.). The Mann–Whitney two‐tailed test was used to assess statistically significant differences.

Article Snippet: Assessment of HSV‐1 load HSV‐1 DNA was detected and quantified in the placental tissues using the HSV‐1 polymerase chain reaction (PCR) kit (GeneProof a.s., Brno, Czech Republic), according to the manufacturer's instructions.

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

Journal: Clinical and Experimental Immunology

Article Title: Enhanced expression of IFI16 and RIG‐I in human third‐trimester placentas following HSV‐1 infection

doi: 10.1111/cei.13143

Figure Lengend Snippet: Analysis of the supernatants concentrations of (a) interferon (IFN)‐β and (b) tumour necrosis factor (TNF)‐α in decidual and chorionic villous tissue after herpes simplex type 1 (HSV‐1) infection. Decidual and chorionic villi explants were infected with 1 × 105 plaque‐forming units (PFU)/ml HSV‐1 (n = 8). Culture supernatants were harvested at 24 and 48 h post‐infection. The boxes indicate the 25th and 75th percentiles, while the bands near the middle indicate the median values. Data are expressed as the mean ± standard error of the mean (s.e.m.). The Mann–Whitney two‐tailed test was used to assess statistically significant differences.

Article Snippet: Assessment of HSV‐1 load HSV‐1 DNA was detected and quantified in the placental tissues using the HSV‐1 polymerase chain reaction (PCR) kit (GeneProof a.s., Brno, Czech Republic), according to the manufacturer's instructions.

Techniques: Infection, MANN-WHITNEY, Two Tailed Test

Journal: Clinical and Experimental Immunology

Article Title: Enhanced expression of IFI16 and RIG‐I in human third‐trimester placentas following HSV‐1 infection

doi: 10.1111/cei.13143

Figure Lengend Snippet: The relative expression of DDX58, IFIH1, IFI16 and TLR3 in mock‐infected and herpes simplex type 1 (HSV‐1)‐infected Vero cells (n = 4 independent experiments; passages 134–135). Quantitative real‐time polymerase chain reaction (qRT–PCR) was performed using YWHAZ as a housekeeping gene. The data are reported as the mean values ± standard error of the mean (s.e.m.). The Mann–Whitney two‐tailed test was used to assess statistically significant differences.

Article Snippet: Assessment of HSV‐1 load HSV‐1 DNA was detected and quantified in the placental tissues using the HSV‐1 polymerase chain reaction (PCR) kit (GeneProof a.s., Brno, Czech Republic), according to the manufacturer's instructions.

Techniques: Expressing, Infection, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, MANN-WHITNEY, Two Tailed Test

Journal: Journal of Clinical Microbiology

Article Title: Laboratory Diagnosis of Neonatal Herpes Simplex Virus Infections

doi: 10.1128/JCM.01460-18

Figure Lengend Snippet: FDA-cleared molecular assays for HSV a

Article Snippet: These nucleic acid amplification-based assays are sensitive and specific, and most can be completed within 1 to 2 h. Compared to virus culture and antigen detection, these molecular methods require instrumentation, and their supply costs are also slightly higher. table ft1 table-wrap mode="anchored" t5 TABLE 2 caption a7 Assay and manufacturer Virus detected and HSV gene target Principle Instrument(s) Intended use Lyra Direct HSV 1 + 2/VZV, Quidel HSV-1 and -2, VZV (amplification targets not specified by manufacturer) Real-time PCR Life Technologies QuantStudio Dx, Applied Biosystems 7500 Fast Dx, Cepheid SmartCycler II System Cutaneous or mucocutaneous lesion samples Solana HSV 1 + 2/VZV, Quidel HSV-1 and -2, VZV (amplification targets not specified by manufacturer) HDA Solana instrument Cutaneous or mucocutaneous lesion samples AmpliVue HSV 1 + 2, Quidel HSV-1 and -2 (amplification target not specified by manufacturer) HDA AmpliVue cassette (disposable and self-contained cartridge) Cutaneous or mucocutaneous lesion specimens illumigene HSV 1&2, Meridian Bioscience HSV-1 and -2, HSV gB LAMP illumipro-10 Cutaneous or mucocutaneous lesion specimens Simplexa HSV 1 & 2 Direct, DiaSorin HSV-1 and -2, HSV DNA polymerase Real-time PCR LIAISON MDX Genital lesion; cutaneous/mucocutaneous swab samples; CSF MultiCode-RTx HSV 1&2, EraGen (currently under Luminex) HSV-1 and -2 HSV gB Real-time PCR Roche LightCycler Vaginal lesion Aries HSV 1&2 assay, Luminex HSV-1 and -2 (amplification target not specified by manufacturer) Real-time PCR Aries system Cutaneous or mucocutaneous specimens FilmArray ME panel, BioFire HSV-1 and -2 plus 12 other pathogens (amplification target not specified by manufacturer) Real-time PCR, multiplex BioFire instrument CSF for meningitis and/or encephalitis Open in a separate window a HSV, herpes simplex virus; VZV, varicella zoster virus; HSV gB, HSV surface glycoprotein B; ME, meningitis/encephalitis; HDA, helicase-dependent isothermal DNA amplification; LAMP, loop-mediated isothermal DNA amplification.

Techniques: Amplification, Real-time Polymerase Chain Reaction, Luminex, Multiplex Assay

Journal: Journal of Clinical Virology

Article Title: Amplification chemistries in clinical virology

doi: 10.1016/j.jcv.2019.03.015

Figure Lengend Snippet: Commercially available nucleic acid amplification assays for viral pathogens a .

Article Snippet: Sentosa SA201 HSV-1/2 PCR Test , Vela Diagnostics , Real-time PCR , HSV 1, HSV 2.

Techniques: Amplification, Real-time Polymerase Chain Reaction, HPV Assay, Multiplex Assay, Viral-load Assay, Quantitative RT-PCR, Virus, Luminex, Reverse Transcription Polymerase Chain Reaction, Microarray, Aptima HCV Quant Dx Assay, CytoScan DX Assay, DNA Amplification

Journal: Journal of Clinical Microbiology

Article Title: Laboratory Diagnosis of Neonatal Herpes Simplex Virus Infections

doi: 10.1128/JCM.01460-18

Figure Lengend Snippet: FDA-cleared molecular assays for HSV a

Article Snippet: These nucleic acid amplification-based assays are sensitive and specific, and most can be completed within 1 to 2 h. Compared to virus culture and antigen detection, these molecular methods require instrumentation, and their supply costs are also slightly higher. table ft1 table-wrap mode="anchored" t5 TABLE 2 caption a7 Assay and manufacturer Virus detected and HSV gene target Principle Instrument(s) Intended use Lyra Direct HSV 1 + 2/VZV, Quidel HSV-1 and -2, VZV (amplification targets not specified by manufacturer) Real-time PCR Life Technologies QuantStudio Dx, Applied Biosystems 7500 Fast Dx, Cepheid SmartCycler II System Cutaneous or mucocutaneous lesion samples Solana HSV 1 + 2/VZV, Quidel HSV-1 and -2, VZV (amplification targets not specified by manufacturer) HDA Solana instrument Cutaneous or mucocutaneous lesion samples AmpliVue HSV 1 + 2, Quidel HSV-1 and -2 (amplification target not specified by manufacturer) HDA AmpliVue cassette (disposable and self-contained cartridge) Cutaneous or mucocutaneous lesion specimens illumigene HSV 1&2, Meridian Bioscience HSV-1 and -2, HSV gB LAMP illumipro-10 Cutaneous or mucocutaneous lesion specimens Simplexa HSV 1 & 2 Direct, DiaSorin HSV-1 and -2, HSV DNA polymerase Real-time PCR LIAISON MDX Genital lesion; cutaneous/mucocutaneous swab samples; CSF MultiCode-RTx HSV 1&2, EraGen (currently under Luminex) HSV-1 and -2 HSV gB Real-time PCR Roche LightCycler Vaginal lesion Aries HSV 1&2 assay, Luminex HSV-1 and -2 (amplification target not specified by manufacturer) Real-time PCR Aries system Cutaneous or mucocutaneous specimens FilmArray ME panel, BioFire HSV-1 and -2 plus 12 other pathogens (amplification target not specified by manufacturer) Real-time PCR, multiplex BioFire instrument CSF for meningitis and/or encephalitis Open in a separate window a HSV, herpes simplex virus; VZV, varicella zoster virus; HSV gB, HSV surface glycoprotein B; ME, meningitis/encephalitis; HDA, helicase-dependent isothermal DNA amplification; LAMP, loop-mediated isothermal DNA amplification.

Techniques: Virus, Amplification, Real-time Polymerase Chain Reaction, Luminex, Multiplex Assay