Polymerase chain reaction-based ganciclovir resistance testing of ocular fluids for cytomegalovirus retinitis.
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ABSTRACT:
SUBMITTER: Yeh S
PROVIDER: S-EPMC4064592 | biostudies-literature | 2012 Jan
REPOSITORIES: biostudies-literature
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Polymerase chain reaction-based ganciclovir resistance testing of ocular fluids for cytomegalovirus retinitis.
Yeh Steven S Fahle Gary G Forooghian Farzin F Faia Lisa J LJ Weichel Eric D ED Stout J Timothy JT Flaxel Christina J CJ Lauer Andreas K AK Sen H Nida HN Nussenblatt Robert B RB
Archives of ophthalmology (Chicago, Ill. : 1960) 20120101 1
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Project description:Ganciclovir (GCV) is one of the most widely used antiviral drugs for the treatment of cytomegalovirus (CMV) retinitis. In this context, the aim of this study was to design in situ thermosensitive hydrogels for GCV ocular delivery by intravitreal injection to achieve sustained drug release behavior and improved ocular bioavailability in the treatment of CMV retinitis. A thermosensitive poly-(β-butyrolactone-co-lactic acid)-polyethylene glycol-poly (β-butyrolactone-co-lactic acid) (PBLA-PEG-PBLA) triblock copolymer was synthesized by ring-opening polymerization and characterization. The GCV-loaded PBLA-PEG-PBLA in situ hydrogels (15%, w/w) were then prepared with drug concentration at 2 mg·mL-1 and the gelation temperatures, rheological properties, in vitro degradation and syringeability of in situ hydrogels for intravitreal injection were also investigated. Membraneless dissolution model was used to explore drug release behavior of PBLA-PEG-PBLA in situ hydrogel. The results indicated that more than 45 and 85% of GCV can be released within 24 and 96 h, respectively, which was verified by a non-Fickian diffusion mechanism. In vivo ocular pharmacokinetics study showed that area under drug-time curve (AUC) and half-life of PBLA-PEG-PBLA in situ hydrogel was higher (AUC was 61.80 μg·mL-1·h (p < .01) and t1/2 was 10.29 h in aqueous humor; AUC was 1008.66 μg·mL-1·h (p < .01) and t1/2 was 13.26 h (p < .01) in vitreous) than GCV injection with extended therapeutic activity. Based on obtained results, it was concluded that the thermosenstive PBLA-PEG-PBLA in situ hydrogel is a promising carrier of GCV for intravitreal injection.
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Project description:PurposeTo associate detection of potential pathogen DNA in endophthalmitis with clinical outcomes.DesignProspective cohort study.MethodsPatients in whom endophthalmitis was diagnosed following an intraocular procedure were recruited. Clinical outcome data from baseline, week-1, month-1, and month-3 visits were collected. Intraocular biopsy samples were cultured by standard methods. Quantitative polymerase chain reaction (qPCR) was performed for specific pathogens and whole-genome sequencing (WGS).ResultsA total of 50 patients (mean age 72 years old; 52% male) were enrolled. Twenty-four cases were culture-positive and 26 were culture-negative. WGS identified the cultured organism in 76% of culture-positive cases and identified potential pathogens in 33% of culture-negative cases. Month-1 and -3 visual acuities did not vary by pathogen-positive versus pathogen-negative cases as detected by either culture or WGS. Visual outcomes of Staphylococcus epidermidis endophthalmitis were no different than those of pathogen-negative cases, whereas the patients infected with other pathogens showed worse outcome. Higher baseline bacterial DNA loads of bacteria other than those of S epidermidis detected by WGS were associated with worse month-1 and -3 visual acuity, whereas the S epidermidis loads did not appear to influence outcomes. Torque teno virus (TTV) and Merkel cell polyomavirus (MCV) were detected by qPCR in 49% and 19% of cases, respectively. Presence of TTV at presentation was associated with a higher rate of secondary pars plana vitrectomy (P = .009) and retinal detachment (P = .022).ConclusionsThe presence and higher load of bacteria other than S epidermidis detected by WGS or DNA from TTV by qPCR in ocular fluids is associated with worse outcomes in post-procedure endophthalmitis.
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Project description:Interventions: Patients with the detection of either cytomegalic cells or IHC-CMV cells were diagnosed with CMV colitis.,Patients older than 18 years with clinical suspicion of CMV colitis but negative biopsy for CMV.,Asymptomatic patients underwent colonoscopy for screening.;Diagnostic,Diagnostic,Diagnostic;CMV colitis,Non-CMV colitis ,Healthy volunteers
Primary outcome(s): Diagnostic performance After colonoscopy Sensitivity, Specificity
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Project description:Purpose:Cytomegalovirus (CMV) has been reported to cause anterior uveitis in the immunocompetent people. Recurrence of this viral uveitis poses a mangement dilemma. With real-time polymerase chain reaction(PCR), it is possible to confirm the clinical diagnosis. We report a case of recurrent CMV anterior uveitis documented by real-time PCR. Observations:A 42 year old man developed PCR proven CMV anterior uveitis. It resolved with oral and topical antivirals but recurred after ten months. The recurrence was controlled by restarting the oral antivirals. Real-time PCR was used to sequentially document the inital infection, the subsequent resolution and the recurrence of the infection. Conclusions and Importance:Real-time PCR is a useful tool in the management of CMV anterior uveitis.
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Project description:BackgroundCongenital cytomegalovirus (CMV) infection is an important cause of hearing loss, and most infants at risk for CMV-associated hearing loss are not identified early in life because of failure to test for the infection. The standard assay for newborn CMV screening is rapid culture performed on saliva specimens obtained at birth, but this assay cannot be automated. Two alternatives--real-time polymerase-chain-reaction (PCR)-based testing of a liquid-saliva or dried-saliva specimen obtained at birth--have been developed.MethodsIn our prospective, multicenter screening study of newborns, we compared real-time PCR assays of liquid-saliva and dried-saliva specimens with rapid culture of saliva specimens obtained at birth.ResultsA total of 177 of 34,989 infants (0.5%; 95% confidence interval [CI], 0.4 to 0.6) were positive for CMV, according to at least one of the three methods. Of 17,662 newborns screened with the use of the liquid-saliva PCR assay, 17,569 were negative for CMV, and the remaining 85 infants (0.5%; 95% CI, 0.4 to 0.6) had positive results on both culture and PCR assay. The sensitivity and specificity of the liquid-saliva PCR assay were 100% (95% CI, 95.8 to 100) and 99.9% (95% CI, 99.9 to 100), respectively, and the positive and negative predictive values were 91.4% (95% CI, 83.8 to 96.2) and 100% (95% CI, 99.9 to 100), respectively. Of 17,327 newborns screened by means of the dried-saliva PCR assay, 74 were positive for CMV, whereas 76 (0.4%; 95% CI, 0.3 to 0.5) were found to be CMV-positive on rapid culture. Sensitivity and specificity of the dried-saliva PCR assay were 97.4% (95% CI, 90.8 to 99.7) and 99.9% (95% CI, 99.9 to 100), respectively. The positive and negative predictive values were 90.2% (95% CI, 81.7 to 95.7) and 99.9% (95% CI, 99.9 to 100), respectively.ConclusionsReal-time PCR assays of both liquid- and dried-saliva specimens showed high sensitivity and specificity for detecting CMV infection and should be considered potential screening tools for CMV in newborns. (Funded by the National Institute on Deafness and Other Communication Disorders.).
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Project description:BackgroundThe purpose of this study is to examine the clinical outcomes achieved by using initial high-dose intravitreal ganciclovir injections to treat cytomegalovirus retinitis in patients without human immunodeficiency virus (HIV) infection.MethodsTwenty-four eyes (24 patients) with cytomegalovirus retinitis received multiple intravitreal injections of ganciclovir in weekly intervals. A higher dose (6 mg) of ganciclovir was applied at the first intravitreal injection, and a lower dose was used for maintenance. Anterior aqueous humour was obtained before each injection. The best-corrected visual acuity and cytomegalovirus loads in the anterior aqueous humour were measured.ResultsThe mean cytomegalovirus DNA load in aqueous humour decreased significantly from (2.59 ± 2.28) × 105 copies/mL at baseline to (1 ± 1.76) × 104 copies/mL one month later. The aqueous cytomegalovirus DNA load was negative in 17 eyes (70.8%) one month later. No obvious improvement of best-corrected visual acuity was found during the treatment. A positive correlation was proven between initial cytomegalovirus DNA titers in aqueous humour and the total number of intravitreal injections of ganciclovir, as well as between the baseline and final best-corrected visual acuities. No severe complications developed.ConclusionsAn initial high dose of ganciclovir (6 mg) and continuous intravitreal injections of ganciclovir could significantly decrease the cytomegalovirus load in HIV-negative patients with cytomegalovirus retinitis.Trial registrationhttp://clinicaltrials.gov, NCT03598452, retrospectively registered on 24 July 2018.
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