Original articleDetection and genotyping of herpes simplex virus types 1 and 2 by polymerase chain reaction
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2015, Life SciencesCitation Excerpt :Cycling conditions were as follows: denaturation of DNA template at 95 °C for 3 min, followed by 35 cycles of 94 °C for 30 s, 56 °C for 30 s, 72 °C for 30 s and a final extension step at 72 °C for 8 min. All b-globin-positive samples were tested for HPV, HSV-1 and 2, and CT DNA, using previously described specific primers [24–26] (Metabion, Martinsried, Germany) listed in Table 1. HPV PCR amplification was carried out in 50 μL reaction mixture with 1 × PCR Buffer, 2 mM MgCl2, 0.20 mM dNTPs, 12.5 pmol of each primer, 1 unit Taq polymerase and 50–100 ng DNA template.
Herpes simplex virus type 1 is the main cause of genital herpes in women of Natal, Brazil
2012, European Journal of Obstetrics and Gynecology and Reproductive BiologyCitation Excerpt :The conditions for both reactions were as follows: incubation at 50 °C for 2 min, denaturation of DNA at 95 °C for 5 min, followed by 40 cycles of 94 °C for 1 min, 45 s at 58 °C for annealing, an extension step at 72 °C for 30 s, and a final extension step at 72 °C for 10 min. The primers HSV-1a (5′-CCCTGTCTCGCGCGAGCCAC-3′) and HSV-1b (5′-TCAGCCACCCATACGCGTAA-3′), which amplify a fragment of 142 bp [19], were used to detect DNA from HSV-1, while the primers HSV-2 (A) (5′-GGACGAGGCGCCAAAGCACACG-3′) and HSV-2 (B) (5′-TCCGTCCAGTCGTTTATCTTCAC-3′), which generate a product of 270 bp [20], were used for HSV-2. The products of PCRs underwent vertical electrophoresis on polyacrylamide gel at 8% [16], with subsequent revelation by silver [18].
Simultaneous detection of herpes simplex virus types 1 and 2 by real-time PCR and Pyrosequencing
2005, Journal of Clinical VirologyA duplex PCR assay for detection and genotyping of Herpes simplex virus in cerebrospinal fluid
1999, Molecular and Cellular ProbesQuantitation of herpes simplex viral DNA in vero cells for evaluation of an antiviral agent using the polymerase chain reaction
1998, Journal of Virological Methods