Aqueous humor polymerase chain reaction in uveitis – utility and safety

Chronopoulos, Argyrios; Roquelaure, Daniel; Souteyrand, Georges; Seebach, Jörg Dieter; Schutz, James Scott; Thumann, Gabriele

doi:10.1186/s12886-016-0369-z

Cited by 55 publications

(46 citation statements)

References 39 publications

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“…The qPCR assay revised from previous publications was established (Table ) . All the qPCR detection was performed according to the manufacturer's instructions on a Roche LightCycler 480 system.…”

Section: Methodsmentioning

confidence: 99%

“…The ddPCR detection system revised from previous publications is described as follows . The ddPCR detection system was performed using ddPCR Supermix for Probes kit (no dUTP,Bio‐Rad) on a QX200 Droplet Digital PCR System according to manufacturer's manual.…”

Section: Methodsmentioning

confidence: 99%

“…Several factors have been postulated as the contributors to PSS development, including abnormal vascular processes, autonomic defect, allergic condition and variation of developmental glaucoma. Recently, it was reported that PSS is related to viral infections such as human cytomegalovirus (HCMV), herpes simplex virus (HSV) and varicella zoster virus (VZV) . As an elusive disease, new insights into possible infectious aetiologies of PSS may lead to the development of effective therapeutic strategies.…”

Section: Introductionmentioning

confidence: 99%

“…With the characteristics of rapid analysis, conventional polymerase chanin reaction (PCR) has been useful for the specific diagnosis of infectious aetiology in the aqueous humour and efficient treatment of uveitis patients . However, because of the diversity of conventional PCR (quantitative real‐time polymerase chain reaction, qPCR) platforms used in different clinical laboratories, there may be large differences on commercial or in‐house calibration materials PCR components, etc., which in turn result in significant variability of the reported quantitative and qualitative data among different laboratories .…”

Section: Introductionmentioning

confidence: 99%

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Digital droplet polymerase chain reaction analysis of common viruses in the aqueous humour of patients with Posner‐Schlossman syndrome in Chinese population

Cao

Tan

Zhang

et al. 2018

Clinical Exper Ophthalmology

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Background To compare the detection results consistency of quantitative polymerase chain reaction (qPCR) and digital droplet polymerase chain reaction (ddPCR), and determine the value of ddPCR for viral detection in the aqueous humour. Methods A total of 130 aqueous humour samples were collected, including 60 patients with Posner‐Schlossman syndrome (PSS) in case group and 70 elderly patients with senile cataract in control group. The target nucleic acid fragments of human cytomegalovirus (HCMV), herpes simplex virus, Epstein‐Barr virus and varicella zoster virus in aqueous humour were analysed by qPCR and ddPCR, respectively, for the diagnosis and curative effect monitoring of pathogen‐induced PSS. Samples with inconsistent results were verified by next‐generation sequencing. Results There were 27 and 20 HCMV‐positive cases detected in the case group by ddPCR and qPCR, respectively. ddPCR increased the sensitivity for the HCMV virus detection from 400 to 100 copies/mL. No other pathogens were found in this study. The results of ddPCR were consistent with that of next generation sequencing. The mean (SD) of Lg (HCMV copies/mL) detected by ddPCR and qPCR were 1.66 (1.92) and 1.10 (1.61), respectively (P < 0.001). Compared with qPCR, results of ddPCR showed better consistency with validity of clinical treatment. All patients with ddPCR‐positive results had good validity on antiviral therapy, exhibiting anterior chamber inflammation remission, resolution of corneal oedema and good IOP control within 1 month. Conclusions HCMV was the leading cause of pathogen‐induced PSS in the Chinese population. ddPCR was a promising tool for early detection, accurate diagnosis and therapeutic validity monitoring of pathogen‐induced PSS. The high sensitivity of ddPCR could avoid repeated anterior chamber tap.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Digital droplet polymerase chain reaction analysis of common viruses in the aqueous humour of patients with Posner‐Schlossman syndrome in Chinese population

Cao

Tan

Zhang

et al. 2018

Clinical Exper Ophthalmology

View full text Add to dashboard Cite

show abstract

“…Emperical treatment with corticosteroids and immunosuppressives should not be given in all patients with PU. Ancillary tests 20 and aqueous humor analysis 21 helps in establishing the diagnosis. Follow up is important to look for recurrence and complications because early diagnosis and correct treatment can prevent visual loss.…”

Section: Discussionmentioning

confidence: 99%

A clinical profile of posterior uveitis in a multispecialty hospital

Annamalai¹,

J²,

Muthukumar³

2017

Int j curr adv res

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Potential Diagnosis of Vitreoretinal Lymphoma by Detection ofMYD88Mutation in Aqueous Humor With Ultrasensitive Droplet Digital Polymerase Chain Reaction

et al. 2018

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IMPORTANCEThe diagnostic workup of patients suspected of having vitreoretinal lymphoma (VRL) is primarily based on vitreous fluid analysis, including the recently emerging myeloid differentiation primary response gene 88 (MYD88) mutation analysis. Aqueous humor paracentesis is a relatively less invasive and safer procedure than taking vitreous fluid specimens, and aqueous humor-based MYD88 mutation analysis would provide an additional liquid biopsy tool to diagnose and monitor patients with VRL. OBJECTIVE To investigate whether the detection of MYD88 L265P by highly sensitive droplet digital polymerase chain reaction (ddPCR) is feasible in the vitreous fluid and aqueous humor of patients with VRL. DESIGN, SETTING, AND PARTICIPANTSThis cohort study includes aqueous humor and vitreous fluid samples from patients with VRL who were treated at the University Medical Center Utrecht, in Utrecht, the Netherlands, from August 2005 to August 2017. Ocular fluids were randomized and masked before MYD88 L265P analysis, which was performed using an in-house validated ddPCR platform. Patients with uveitis were included as a comparison group. MAIN OUTCOMES AND MEASURESThe presence of MYD88 L265P mutation detected by ddPCR in AH and VF. RESULTSThe study included 96 samples from 63 individuals, including 23 patients with VRL (of whom 10 were female and 13 male, with a mean [SD] age of 72 [7.3] years) and 40 individuals with uveitis (of whom 23 were female and 17 male, with a mean [SD] age of 58 [20.9] years). In 17 of 23 patients with VRL (74%), MYD88 L265P was detected; it was not detected in any of the patients with uveitis. It was detectable in both vitreous fluid and aqueous humor samples. In the paired samples, the mutation was detected in 8 of 9 aqueous humor samples (89%) of the MYD88 L265P-positive vitreous fluid samples. In vitreous fluid, the MYD88 ddPCR test showed a sensitivity of 75% (95% CI, 50%-92%) and a positive predictive value of 100%; in aqueous humor, sensitivity was 67% (95% CI, 42%-92%), and positive predictive value was 100%. Specificity was 100% in both fluids. After treatment, the mutation was no longer detectable in any ocular fluids. CONCLUSIONS AND RELEVANCEThe high concordance between aqueous humor and vitreous fluid samples suggests that use of the easily accessible aqueous humor is nearly as informative as vitreous fluid in the identification of key somatic mutations in patients with VRL. This approach may provide an additional minimally invasive tool for accurate diagnosis, detection of recurrence, and monitoring of treatment.

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Aqueous humor polymerase chain reaction in uveitis – utility and safety

Cited by 55 publications

References 39 publications

Digital droplet polymerase chain reaction analysis of common viruses in the aqueous humour of patients with Posner‐Schlossman syndrome in Chinese population

Digital droplet polymerase chain reaction analysis of common viruses in the aqueous humour of patients with Posner‐Schlossman syndrome in Chinese population

A clinical profile of posterior uveitis in a multispecialty hospital

Potential Diagnosis of Vitreoretinal Lymphoma by Detection ofMYD88Mutation in Aqueous Humor With Ultrasensitive Droplet Digital Polymerase Chain Reaction

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