Diagnosis of ocular toxoplasmosis by two polymerase chain reaction (PCR) examinations: qualitative multiplex and quantitative real-time

Sugita, Sunao; Ogawa, Manabu; Inoue, Shizu; Shimizu, Norio; Mochizuki, Manabu

doi:10.1007/s10384-011-0065-8

Cited by 33 publications

(26 citation statements)

References 20 publications

(38 reference statements)

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“…PCR for the detection of a Toxoplasma-specific gene region is reported to be useful [13,127]. In our previous study [13], PCR analysis using a combination of multiplex qualitative PCR and real-time quantitative PCR assays of intraocular fluids, mostly the aqueous humor, detected DNA of Toxoplasma gondii in all patients with active uveitis, but not in patients with inactive old lesions (Fig.…”

Section: Toxoplasma Gondiimentioning

confidence: 92%

“…Multiplex PCR has the advantages of combining several different primer pairs in the same amplification reaction, with the net result of producing different specific infectious amplicons in ocular inflammatory diseases. This technology facilitates the comprehensive measurements of the genomes of many pathogenic agents [9][10][11][12][13][14]. Therefore, specific genomic DNA from different viruses, bacteria, and fungi can be measured simultaneously in a small sample volume, which is particularly important for intraocular fluid samples, such as aqueous humor.…”

Section: Multiplex Pcr and Real-time Pcrmentioning

confidence: 99%

“…The multiplex PCR was designed to detect genomic DNA of all eight human herpes viruses (HHVs): HHV1 (herpes simplex virus type 1; HSV1), HHV2, HHV3 (varicella-zoster virus; VZV), HHV4 (Epstein-Barr virus; EBV), HHV5 (cytomegalovirus; CMV), HHV6, HHV7, and HHV8 [11,12]. The PCR assay was also designed to qualitatively detect genomic DNA of toxoplasma (Toxoplasma gondii B1 gene) [13], parvovirus B19 (Parvo B19), BK virus (BKV), and JC virus (JCV). Parvo B19, BKV, and JCV genomic DNA are often detected in intraocular fluid samples from patients with ocular inflammatory diseases, especially immunodeficient patients (unpublished observation).…”

Section: A Comprehensive Pcr Systemmentioning

confidence: 99%

“…In our laboratory, this PCR assay is performed with a LightCycler PCR System (Roche Life Sciences). The multiplex PCR assay screens for the infectious genomes of all human herpes viruses (HHV1-8) [11,12] and ocular toxoplasma [13]. When positive results are generated by the multiplex PCR, then quantitative real-time PCR is performed to measure the copy number of the genome in the sample and to confirm its pathogenic role.…”

Section: A Comprehensive Pcr Systemmentioning

confidence: 99%

“…Although the molecular diagnosis of Toxocara larva has been reported by using liver biopsy specimens [131], experimental animal models of ocular toxocariasis [132], or carnivore feces [133], PCR assays are less frequently used to diagnose ocular toxocariasis using results of the multiplex PCR using the aqueous humor of the eye were positive for Toxoplasma gondii DNA, and real-time PCR revealed 1.1 9 10 6 copies/ml. Reproduced from Sugita et al [13] with permission Fig. 12 Ocular toxocariasis.…”

Section: Toxocara Canis and T Catimentioning

confidence: 99%

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A new era of uveitis: impact of polymerase chain reaction in intraocular inflammatory diseases

et al. 2016

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Uveitis is a sight-threatening intraocular inflammatory disorder which may occur from both infectious and non-infectious or autoimmune causes. The frequency of infectious uveitis and autoimmune uveitis varies depending on countries and regions. According to a nationwide survey conducted by the Japanese Ocular Inflammation Society, infectious and non-infectious uveitis accounted for 16.4 and 50.1% of new patients, respectively while the remaining 33.5% of new uveitis cases were not classified or were idiopathic uveitis. Infectious uveitis is particularly important because it causes tissue damage to the eye and may result in blindness unless treated. However, it can be treated if the pathogenic microorganisms are identified promptly and accurately. Remarkable advancements in molecular and immunological technologies have been made in the last decade, and the diagnosis of infectious uveitis has been greatly improved by the application of molecular and immunological investigations, particularly polymerase chain reaction (PCR). PCR performed on a small amount of ocular samples provides a prompt, sensitive, and specific molecular diagnosis of pathogenic microorganisms in the eye. This technology has opened a new era in the diagnosis and treatment of uveitis, enabling physicians to establish new clinical entities of uveitis caused by infectious microorganisms, identify pathogens in the eyes of many patients with uveitis, and determine prompt diagnosis and appropriate therapy. Here we review the PCR process, new PCR tests specialized for ocular diseases, microorganisms detected by the PCR tests, diseases in the eye caused by these microorganisms, and the clinical characteristics, diagnosis, and therapy of uveitis.

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Section: Toxoplasma Gondiimentioning

confidence: 92%

Section: Multiplex Pcr and Real-time Pcrmentioning

confidence: 99%

Section: A Comprehensive Pcr Systemmentioning

confidence: 99%

Section: A Comprehensive Pcr Systemmentioning

confidence: 99%

Section: Toxocara Canis and T Catimentioning

confidence: 99%

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A new era of uveitis: impact of polymerase chain reaction in intraocular inflammatory diseases

et al. 2016

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Frequency of Toxoplasma gondii in the retina in eye banks in Brazil

et al. 2017

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Ocular toxoplasmosis is the main cause of posterior uveitis worldwide frequently leading to vision loss. In Brazil, the seroprevalence of Toxoplasma gondii infection ranges from 50 to 80% depending of the region studied. The frequency of toxoplasmic retinal scar may reach 18% of the adults in the South of Brazil. Our goal was to determine the frequency of T. gondii DNA in retinas from eye banks from different regions in Brazil. A total of 162 eyes were obtained from eye banks in Manaus (n = 60), Sao Paulo (n = 60), Chapeco (n = 26), and Joinville (n = 16). The retinas were macroscopically analyzed and collected for DNA extraction. Real-time PCR (qPCR) was performed using the T. gondii B1 marker. By qPCR, a higher frequency of T. gondii DNA in the retinas from the eye bank of Joinville (25%) was found when compared to Manaus (5%). The retinas from Sao Paulo and Chapeco were qPCR negative. Clinical examination determined the retina lesions to be compatible with toxoplasmosis in the following frequencies: Joinville (62.5%), Manaus (10%), Sao Paulo (6.7%), and Chapeco (15.4%).

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Development and validation of new diagnostic criteria for acute retinal necrosis

et al. 2014

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Diagnosis of ocular toxoplasmosis by two polymerase chain reaction (PCR) examinations: qualitative multiplex and quantitative real-time

Abstract: The two-step PCR examination to detect toxoplasma DNA is a useful tool for diagnosing ocular toxoplasmosis.

Cited by 33 publications

References 20 publications

A new era of uveitis: impact of polymerase chain reaction in intraocular inflammatory diseases

A new era of uveitis: impact of polymerase chain reaction in intraocular inflammatory diseases

Frequency of Toxoplasma gondii in the retina in eye banks in Brazil

Development and validation of new diagnostic criteria for acute retinal necrosis

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