LOW-VOLUME MULTIPLEX PCR FOR ETIOLOGICAL DIAGNOSIS OF INFECTIOUS UVEITIS
Purpose
To analyse the usefulness of low-volume direct multiplex PCR of intraocular fluid for the etiological diagnosis of uveitis.
Methods
Samples (aqueous humor, n=33; vitreous humor, n=2) from 35 patients with active uveitis (≥2+ cells in anterior chamber (AC)) (July,21 to Sept,22). 20µl of sample was analysed using a direct multiplex qualitative polymerase chain reaction (PCR) assay (Nihon Techno Service Co, Japan, for research purpose) including herpes simplex virus (HSV) 1 and 2; varicella-zoster virus, cytomegalovirus, Epstein-Barr virus (EBV), human herpes virus 6 (HHV6), human T-lymphotropic virus, Treponema pallidum and Toxoplasma gondii).
Results
Uveitis were anterior, intermediate, posterior and panuveitis in 16 (45.7%), 1 (2.8%), 15 (42.8%) and 3 (8.7%) patients, respectively. Overall positivity was 25.7% (9/35 samples); among acute uveitis was 34.8% (8/23 samples); among infectious uveitis (clinical diagnosis) was 39.1% (9/23 samples). Herpes virus was detected in 4 samples: 2 for HHV6 (anterior uveitis), 1 for HSV2 (acute retinal necrosis) and 1 for EBV (neuroretinitis); T. gondii was detected in 4 samples (chorioretinitis) and T. pallidum in another sample (diffuse uveitis). The strip PCR changed the etiological diagnosis in two cases (5.7%; herpetic uveitis or syphilis to toxoplasmosis) and showed a unexpected herpesvirus as causative agent in another two cases. All 12 non-infectious uveitis samples were negative.
Conclusion
For uveitis etiological diagnosis, direct strip PCR was able to demonstrate the infectious agent in one third of our sample with the unique characteristics of using very small sample volume, of testing for multiple pathogens all together and for a rapid results.
Conflict of interest
No
Authors 1
Last name
YAMAMOTO
Initials of first name(s)
JH
Department
Ophthalmology, LIM 33, Faculdade de Medicina, Universidade de Sao Paulo
City
Sao Paulo
Country
Brazil
Authors 2
Last name
Ferracioli-Oda
Initials of first name(s)
E
Department
Ophthalmology, Faculdade de Medicina, Universidade de Sao Paulo
City
Sao Paulo
Country
Brazil
Authors 3
Last name
Gouveia
Initials of first name(s)
MGS
Department
Clinical and Experimental Gastroenterology Laboratory, LIM 07, FMUSP
City
Sao Paulo
Country
Brazil
Authors 4
Last name
Pinho
Initials of first name(s)
JR
Department
Clinical and Experimental Gastroenterology Laboratory, LIM 07, FMUSP
City
Sao Paulo
Country
Brazil
Authors 5
Last name
Tanaka
Initials of first name(s)
T
Department
Ophthalmology, Faculdade de Medicina, Universidade de Sao Paulo
City
Sao Paulo
Country
Brazil
Authors 6
Last name
Coelho
Initials of first name(s)
V
Department
Laboratory of Immunology, Heart Institute, FMUSP
City
Sao Paulo
Country
Brazil
Authors 7
Last name
Bispo
Initials of first name(s)
PJ
Department
Ophthalmology, Harvard Medical School
City
Boston
Country
United States
Authors 8
Last name
Hirata
Initials of first name(s)
CE
Department
Ophthalmology, Faculdade de Medicina, Universidade de Sao Paulo
City
Sao Paulo
Country
Brazil
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