Development of a Nested Polymerase Chain Reaction Assay for the Detection and Identification of Pythium insidiosum

Grooters, Amy M.; Gee, Melaney K.

doi:10.1892/0891-6640(2002)016<0147:doanpc>2.3.co;2

Cited by 78 publications

(74 citation statements)

References 24 publications

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“…This was based on the gross lesions, histopathology and identification of the pathogen or nested-PCR with specific primers, as proposed by Grooters and Gee (2002). Concerning the macroscopic aspects, the granulomatous and ulcerative pattern, observation of pruritis, presence of serosanguineous to mucopurulent exudates and presence of fistulous tracts with kunkers were considered.…”

Section: Diagnosismentioning

confidence: 99%

Equine pythiosis: Report of 28 cases from São Paulo State, Brazil

et al. 2015

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Pythiosis is a granulomatous lesion of the skin and subcutaneous tissues caused by Pythium insidiosum, a microorganism belonging to the Stramenopila Kingdom and the Oomycetes Class. The pathogen is commonly found in water environments, mainly in tropical areas of the world. Twenty eight cases of equine pythiosis were presented at the Large Animal Surgery Department of the School of Veterinary Medicine and Animal Science, São Paulo State University, Brazil in six years. Among the 28 animals, 13 presented distal lesions on the limbs, with four being immediately euthanized and seven dying due to loss of body condition leading to cachexia. The horses presented with one or more wounds in the body, distributed as follows: limbs (13 distal, four proximal), abdominal region (7), thoracic region (1), pectoral region (1), lumbar region (1), nasal region (1) and prepuce (1). The diagnosis was made by the association of the macroscopic aspects of the lesions with the histopathology, isolation of the pathogen and/or nested-PCR. Treatments included surgical removal of the lesion (16), potassium iodide 67 mg kg -1 PO sid (23), intravenous regional perfusion with 50 mg of amphotericin B (6) and immunotherapy (8). Pythiosis is a disease that develops quickly, but its diagnosis is time-consuming; therefore, establishing an early treatment with special attention to the involvement of distal regions of the limbs is important. Key words: Equine, granulomatous, pythiosis, Pythium insidiosum, surgery, wound ResumoA pitiose é uma doença granulomatosa do tecido cutâneo e subcutâneo causado pelo oomiceto Pythium insidiosum, microorganismo pertencente ao reino Stramenopila, classe Oomycetes. O agente é comumente encontrado em ambientes aquáticos, principalmente nas áreas tropicais do mundo. Foram atendidos 28 casos no Serviço de Clínica Cirúrgica de Grandes Animais da Faculdade de Medicina Veterinária e Zootecnia, UNESP, Botucatu, São Paulo, Brasil. Dentre os 28 animais, 13 apresentaram lesões distais de membro, sendo quatro imediatamente submetidos à eutanásia e sete morreram devido à perda progressiva de condição corporal levando a caquexia. Os equinos apresentaram uma ou mais lesões, distribuídas da seguinte forma: membros (13 distais e quatro proximais), região abdominal (7), região torácica (1), região peitoral (1), região lombar (1), região nasal (1), prepúcio (1). O diagnóstico foi baseado na associação dos achados macroscópicos, histopatológicos, isolamento do patógeno e/ou nested-PCR. Os tratamentos incluíram: remoção cirúrgica (16), iodeto de potássio 67 mg kg -1 VO Sid (23), perfusão regional com 50 mg de anfotericina B (6) e imunoterapia (8 de rápido desenvolvimento, sendo imprescindível o diagnóstico rápido com especial atenção às lesões localizadas na região distal do membro.

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Section: Diagnosismentioning

confidence: 99%

Equine pythiosis: Report of 28 cases from São Paulo State, Brazil

et al. 2015

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“…Diagnosis by culture identification is time-consuming and requires expertise (17). Serodiagnostic (enzyme-linked immunosorbent assay and immunodiffusion) and PCR-based diagnostic tests have been developed, but diagnostic materials are not widely available (5,9,19,30). Since conventional antifungal drugs are not effective in treating pythiosis, surgery is then the main treatment option.…”

mentioning

confidence: 99%

Identification of a Novel 74-Kilodalton Immunodominant Antigen of Pythium insidiosum Recognized by Sera from Human Patients with Pythiosis

Krajaejun¹,

Kunakorn²,

Pracharktam³

et al. 2006

J Clin Microbiol

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The oomycetous, fungus-like, aquatic organism Pythium insidiosum is the etiologic agent of pythiosis, a life-threatening infectious disease of humans and animals that has been increasingly reported from tropical, subtropical, and temperate countries. Human pythiosis is endemic in Thailand, and most patients present with arteritis, leading to limb amputation and/or death, or cornea ulcer, leading to enucleation. Diagnosis of pythiosis is time-consuming and difficult. Radical surgery is the main treatment for pythiosis because conventional antifungal drugs are ineffective. The aims of this study were to evaluate the use of Western blotting for diagnosis of human pythiosis, to identify specific immunodominant antigens of P. insidiosum, and to increase understanding of humoral immune responses against the pathogen. We performed Western blot analysis on 16 P. insidiosum isolates using 12 pythiosis serum samples. These specimens were derived from human patients with pythiosis who had different forms of infection and lived in different geographic areas throughout Thailand. We have identified a 74-kDa immunodominant antigen in all P. insidiosum isolates tested. The 74-kDa antigen was also recognized by sera from all patients with pythiosis but not by control sera from healthy individuals, patients with thalassemia, and patients with various infectious diseases, indicating that Western blot analysis could facilitate diagnosis of pythiosis. Therefore, the 74-kDa antigen is a potential target for developing rapid serodiagnostic tests as well as a therapeutic vaccine for pythiosis. These advances could lead to early diagnosis and effective treatment, crucial factors for better prognosis for patients with pythiosis.

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“…These spores are phagocytized by macrophages and dispersed through the bloodstream or lymphatic channel to the organ systems and formation of subclinical or clinical granulomatous condition amongst other organs including the GI tract (LinBlache et al, 2011). In young aged, large-breed, male dogs Pythiuminsidiosum may produce pyogenic granulomas in mucosa and submucosa of stomach and different places of the GI tract (Grooters and Gee, 2002). Cryptococcus neoformans cause granulomatous gastritis mimicking carcinoma which is a factor of gastric outlet obstruction (Van der Gaag et al, 1991).…”

Section: Etiopathogenesis Of Gastritis and Peptic Ulcer Diseasesmentioning

confidence: 99%