P1437 Real time PCR for monitoring herpes simplex virus load in tracheal aspirates of intensive care unit patients

2007 ◽  
Vol 29 ◽  
pp. S401
Author(s):  
N. De Vos ◽  
A. Vankeerberghen ◽  
A. Boel ◽  
I. Demeyer ◽  
L. Van Hoovels ◽  
...  
2011 ◽  
Vol 50 (3) ◽  
pp. 948-952 ◽  
Author(s):  
J.-F. Jazeron ◽  
C. Barbe ◽  
E. Frobert ◽  
F. Renois ◽  
D. Talmud ◽  
...  

2019 ◽  
Vol 2019 (10) ◽  
Author(s):  
Karishma Seomangal ◽  
Yasir Bashir ◽  
Michael Boland ◽  
Paul Neary

Abstract We present a case of an unexpected cause of bowel ischemia in an intensive care unit patient with herpes simplex virus encephalitis who required an operation. A 79-year-old lady was being worked up and treated for encephalitis with antibiotics and an antiviral. On Day 13, she developed abdominal pain, and an ultrasound showed cholelithiasis but no cholecystitis; thus conservative treatment was advocated. By Day 18, pain localized to the right iliac fossa, and she had an emergency laparotomy that showed bowel ischemia and perforation of the caecum with the cause being a terminal ileal adhesional band. An extended right hemicolectomy and ileostomy was performed. Patients with significant comorbidities who are intensive care unit-dependent may still have unexpected clinical challenges. We advocate an increased clinical vigilance in this cohort for unexpected life-threatening presentations such as bowel ischemia and more specifically the cause of the bowel ischemia.


1999 ◽  
Vol 37 (6) ◽  
pp. 1941-1947 ◽  
Author(s):  
Alexander J. Ryncarz ◽  
James Goddard ◽  
Anna Wald ◽  
Meei-Li Huang ◽  
Bernard Roizman ◽  
...  

We have developed a high-throughput, semiautomated, quantitative fluorescence-based PCR assay to detect and type herpes simplex virus (HSV) DNA in clinical samples. The detection assay, which uses primers to the type-common region of HSV glycoprotein B (gB), was linear from <10 to 108 copies of HSV DNA/20 μl of sample. Among duplicate samples in reproducibility runs, the assay showed less than 5% variability. We compared the fluorescence-based PCR assay with culture and gel-based liquid hybridization system with 335 genital tract specimens from HSV type 2 (HSV-2)-seropositive persons attending a research clinic and 380 consecutive cerebrospinal fluid (CSF) samples submitted to a diagnostic virology laboratory. Among the 162 culture-positive genital tract specimens, TaqMan PCR was positive for 157 (97%) specimens, whereas the quantitative-competitive PCR was positive for 144 (89%) specimens. Comparisons of the mean titer of HSV DNA detected by the two assays revealed that the mean titer detected by the gel-based system was slightly higher (median, 1 log). These differences in titers were in part related to the fivefold difference in the amount of HSV DNA used in the amplicon standards with the two assays. Among the 380 CSF samples, 42 were positive by both assays, 13 were positive only by the assay with the agarose gel, and 3 were positive only by the assay with the fluorescent probe. To define the subtype of HSV DNA detected in the screening assay, we also designed one set of primers which amplifies the gG regions of both types of HSV and probes which are specific to either HSV-1 (gG1) or HSV-2 (gG2). These probes were labeled with different fluorescent dyes (6-carboxyfluorescein for gG2 and 6-hexachlorofluorescein for gG1) to enable detection in a single PCR. In mixing experiments the probes discriminated the correct subtype in mixtures with up to a 7-log-higher concentration of the opposite subtype. The PCR typing results showed 100% concordance with the results obtained by assays with monoclonal antibodies against HSV-1 or HSV-2. Thus, while the real-time PCR is slightly less sensitive than the gel-based liquid hybridization system, the high throughput, the lack of contamination during processing, the better reproducibility, and the better ability to type the isolates rapidly make the real-time PCR a valuable tool for clinical investigation and diagnosis of HSV infection.


2007 ◽  
Vol 81 (5) ◽  
pp. 549-554 ◽  
Author(s):  
Mami NAGASHIMA ◽  
Kenji SADAMASU ◽  
Takayuki SHINKAI ◽  
Yasuko YOSHIDA ◽  
Sumio YAMADA

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