Detection of
            <i>Balamuthia</i>
            Mitochondrial 16S rRNA Gene DNA in Clinical Specimens by PCR

Yagi, Shigeo; Booton, Gregory C.; Visvesvara, Govinda S.; Schuster, Frederick L.

doi:10.1128/jcm.43.7.3192-3197.2005

Cited by 71 publications

(61 citation statements)

References 21 publications

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“…3B) and negative for Acanthamoeba (26,27). For confirmation, the PCR was done on brain tissue sections scraped from slides following deparaffinization and DNA extraction for amplification and sequencing (28). Two primer sets were used: one for Balamuthia mandrillaris and a second one for Acanthamoeba spp.…”

Section: Case Reportmentioning

confidence: 99%

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Diagnosis of First Case of Balamuthia Amoebic Encephalitis in Portugal by Immunofluorescence and PCR

Tavares

Costa²,

Carpenter³

et al. 2006

J Clin Microbiol

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We report here the first Portuguese case of acute fatal granulomatous encephalitis attributed to Balamuthia mandrillaris, initially thought to be a brain tumor, which had a progressive and fatal outcome. Balamuthia mandrillaris is a free-living amoeba recognized as an uncommon agent of granulomatous encephalitis. Infections have been identified in immunocompromised hosts and in immunocompetent pediatric patients. Balamuthia infections are very rare, with only two reported cases in Europe. The case presented here occurred in a previously healthy boy who died 5 weeks after the onset of the symptoms. No evidence of immunological deficiency was noted, and testing for human immunodeficiency virus antibodies was negative. The symptoms were initially thought to be the result of a tumor, but histopathologic examination showed evidence of amoebic infection. Immunofluorescence staining of brain tissue identified B. mandrillaris as the infectious agent. The diagnosis was confirmed with PCR by detecting Balamuthia DNA in formalin-fixed brain tissue sections. Despite initiation of empirical antimicrobial therapy for balamuthiasis, the patient died 3 weeks after being admitted to the hospital. No source of infection was readily apparent. CASE REPORTAn 8-year-old male was transferred to our hospital because of signs of intracranial hypertension. His past medical history was unremarkable except for an episode of varicella infection a year before. Prior to his illness, he practiced swimming in several swimming pools but had not traveled outside Portugal. Symptoms with sporadic headaches started a few months earlier but had become daily for the 2 weeks prior to admission. His parents noted medial esotropia 2 days before admission, and diplopia developed 1 day before. His headaches were more intense in the morning, he had frequent episodes of vomiting and lethargy, and papilledema was noted upon physical examination.Upon admission he appeared well, with a good general status, and his speech was age appropriate. After unilateral eye occlusion, he had no visual defects. Mycobacterium tuberculosis infection was ruled out from urine and sputum samples, and the tuberculin test (2U, SC) was negative. Human immunodeficiency virus type 1 (HIV-1) and HIV-2 serum antibodies and immunologic studies all showed no changes.Fundoscopy revealed bilateral papilledema and right retinal hemorrhages. Neither alteration of consciousness nor neurological focal deficits were noted. There was no fever. Noncontrast computerized tomography (CT) scanning revealed an expansive hypodense lesion in the right frontal lobe, causing distortion of the middle line and ventricular thinning. With contrast enhancement, a heterogeneous cortical lesion surrounded by a huge area of edema was noted for the first time in the right temporal lobe (Fig. 1A and B). A brain tumor was suspected, and dexamethasone was started because of the clinical symptoms. Upon preoperative magnetic resonance imaging (MRI), the frontal lesion was heterogeneous, with hypodense inner and hyperin...

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Section: Case Reportmentioning

confidence: 99%

“…Unfortunately, serum samples from this case were not available for antibody testing. The PCR technique can also demonstrate Balamuthia DNA in CSF of patients (28). Thus, to be a reliable indicator of balamuthiasis, IFA has to be coupled with H&E and indirect immunofluorescent staining of tissue sections and/or PCR of CSF.…”

Section: Case Reportmentioning

confidence: 99%

Diagnosis of First Case of Balamuthia Amoebic Encephalitis in Portugal by Immunofluorescence and PCR

Tavares

Costa²,

Carpenter³

et al. 2006

J Clin Microbiol

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“…These protocols have been applied to the detection of Acanthamoeba in corneal scrapings from keratitis cases (18,22,27,34), for confirmatory laboratory diagnosis of N. fowleri PAM (7,13), and to detect B. mandrillaris in clinical specimens (42). However, none of the published protocols allows for the simultaneous detection of these parasites in CSF and other clinical specimens.…”

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confidence: 99%

Multiplex Real-Time PCR Assay for Simultaneous Detection of Acanthamoeba spp., Balamuthia mandrillaris , and Naegleria fowleri

et al. 2006

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Infections caused by Naegleria fowleri, Acanthamoeba spp., and Balamuthia mandrillaris occur throughout the world and pose many diagnostic challenges. To date, at least 440 cases of severe central nervous system infections caused by these amebas have been documented worldwide. Rapid and specific identification of these free-living amebas in clinical samples is of crucial importance for efficient case management. We have developed a triplex real-time TaqMan PCR assay that can simultaneously identify Acanthamoeba spp., B. mandrillaris, and N. fowleri in the same PCR vessel. The assay was validated with 22 well-characterized amebic strains harvested from cultures and nine clinical specimens that were previously characterized by in vitro culture and/or immunofluorescence assay. The triplex assay demonstrated high specificity and a rapid test completion time of less than 5 h from the reception of the specimen in the laboratory. This assay was able to detect one single ameba per sample analyzed, as determined with cerebrospinal fluid spiked with diluted cultured amebas. This assay could become useful for fast laboratory diagnostic assessment of amebic infections (caused by free-living amebas) in laboratories with adequate infrastructure to perform real-time PCR testing.

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“…PCR analyses of the CSF and IIF staining of biopsy sections are usually used to diagnose GAE (4,5). Recently, high-sensitivity real-time PCR targeting the RNase P gene has been developed (6).…”

Section: Discussionmentioning

confidence: 99%

Successful Treatment of Granulomatous Amoebic Encephalitis with Combination Antimicrobial Therapy

et al. 2013

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Granulomatous amoebic encephalitis (GAE) is a rare but fatal infection. Due to its nonspecific symptoms and laboratory and neuroradiological findings, it is rarely diagnosed antemortem. We herein present the case of a 72-year-old Japanese woman who was diagnosed with GAE following the detection of a pathogen similar to Balamuthia mandrillaris under a microscopic examination of cerebrospinal fluid sediment and who achieved remission with combination antimicrobial therapy. There are no previous reports of pathogens similar to B. mandrillaris being detected in cerebrospinal fluid antemortem; therefore, this case may be used as a benchmark for further studies.

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Detection of Balamuthia Mitochondrial 16S rRNA Gene DNA in Clinical Specimens by PCR

Cited by 71 publications

References 21 publications

Diagnosis of First Case of Balamuthia Amoebic Encephalitis in Portugal by Immunofluorescence and PCR

Diagnosis of First Case of Balamuthia Amoebic Encephalitis in Portugal by Immunofluorescence and PCR

Multiplex Real-Time PCR Assay for Simultaneous Detection of Acanthamoeba spp., Balamuthia mandrillaris , and Naegleria fowleri

Successful Treatment of Granulomatous Amoebic Encephalitis with Combination Antimicrobial Therapy

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