Detection of
            <i>Sporothrix schenckii</i>
            in Clinical Samples by a Nested PCR Assay

Hu, Sindy; Chung, Wen‐Hung; Hung, Shuen‐Iu; Ho, Hung‐Yao; Wang, Zen-Whe; Chen, Chien‐Hsun; Lu, Shu-Chuan; Kuo, Tseng‐Tong; Hong, Hong‐Shang

doi:10.1128/jcm.41.4.1414-1418.2003

Cited by 74 publications

(80 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The first description of PCR for sporotrichosis' diagnosis was reported in 2001 (9). A diagnostic nested PCR assay targeting the S. schenckii 18S rRNA gene was further evaluated and showed high sensitivity and specificity, indicating that PCR may be clinically useful for diagnosis (8).…”

mentioning

confidence: 99%

Rapid Identification of Sporothrix Species by T3B Fingerprinting

et al. 2012

View full text Add to dashboard Cite

bThis article describes PCR fingerprinting using the universal primer T3B to distinguish among species of the Sporothrix complex, S. brasiliensis, S. globosa, S. mexicana, and S. schenckii. This methodology generated distinct banding patterns, allowing the correct identification of all 35 clinical isolates at the species level, confirmed by partial calmodulin (CAL) gene sequence analyses. This methodology is simple, reliable, rapid, and cheap, making it an ideal routine identification system for clinical mycology laboratories. Sporotrichosis is a globally distributed subcutaneous mycosis with areas of high endemicity (11,21) that is caused by the dimorphic fungus Sporothrix schenckii (22). Sporotrichosis has been regarded as a job-related disease occurring as isolated cases or small outbreaks affecting people exposed to plants or soil (1, 6, 7). Rio de Janeiro State, Brazil, is a region of sporotrichosis hyperendemicity where several human and animal cases have been described since 1998 (5, 23).The diagnosis of sporotrichosis is attained by clinical, epidemiological, and laboratorial data, including culture and analysis of phenotypic characteristics. The first description of PCR for sporotrichosis' diagnosis was reported in 2001 (9). A diagnostic nested PCR assay targeting the S. schenckii 18S rRNA gene was further evaluated and showed high sensitivity and specificity, indicating that PCR may be clinically useful for diagnosis (8).S. schenckii was long considered a single taxon, although great genetic variation within this species has been described (10). By associating phenotypic and genotypic features, Marimon et al. (13) recognized three new species, Sporothrix brasiliensis, Sporothrix globosa, and Sporothrix mexicana, and proposed an identification key for these Sporothrix species. S. globosa has worldwide distribution (12, 18), whereas S. brasiliensis is apparently restricted to Brazil (13) and S. mexicana to Mexican environmental samples (13), although the latter was recently identified in Portugal (3). Additionally, these authors have proposed the promotion of S. schenckii var. luriei to the status of a new species, Sporothrix luriei (14). However, identification based only on phenotypic characteristics is often inconclusive due to phenotypic variability within these species. Therefore, new rapid and reliable identification strategies are necessary (19).Analysis of tRNA intergenic spacers was first used to distinguish Streptococcus species (15). It has also been applied successfully for Candida identification (2,16,24). Here, we evaluate T3B PCR fingerprinting to differentiate clinical Sporothrix strains at the species level in comparison to analysis of partial calmodulin (CAL) gene sequences (19).Thirty-five Sporothrix spp. isolates from the Fungal Culture Collection of IPEC/Fiocruz were included in this study approved by the Ethics Commission of the same institution. Among them, S. brasiliensis type strain CBS 120339 (IPEC16490), S. globosa IPEC27135 (18), S. schenckii IPEC29334 (IOC1226) (19), and S. m...

show abstract

mentioning

confidence: 99%

Rapid Identification of Sporothrix Species by T3B Fingerprinting

et al. 2012

View full text Add to dashboard Cite

show abstract

“…To date, rRNA data have been used to identify species associated with fungal dermatoses (21,29,38,39) and PCR-based diagnostic tests have been developed (15,26,62). Psoriasis, a common dermatosis affecting about 3% of the population in industrialized countries (3), is characterized by erythrosquamous cutaneous lesions associated with abnormal patterns of keratinocyte growth and differentiation (35).…”

mentioning

confidence: 99%

Molecular Analysis of Fungal Microbiota in Samples from Healthy Human Skin and Psoriatic Lesions

Paulino¹,

Tseng²,

2006

View full text Add to dashboard Cite

Psoriasis, a common cutaneous disease of unknown etiology, may be triggered by infections, including those due to fungi. Since the fungal community of human skin is poorly characterized, we aimed to analyze the mycological microbiota in healthy skin and psoriatic lesions. Twenty-five skin samples from five healthy subjects (flexor forearm) and three patients with psoriasis were analyzed using broad-range 18S ribosomal DNA (rDNA) and 5.8S rDNA/internal transcribed spacer 2 (ITS2) Malassezia-specific PCR primers. Broadrange PCR analysis indicated that most organisms resembled Malassezia. Malassezia-specific 5.8S/ITS2 analysis of 1,374 clones identified five species and four unknown phylotypes, potentially representing new species. The species distribution appears largely host specific and conserved in different sites of healthy skin. In three subjects, the Malassezia microbiota composition appeared relatively stable over time. Samples of Malassezia microbiota from healthy skin and psoriatic lesions were similar in one patient but substantially different in two others. These data indicate the predominance of Malassezia organisms in healthy human skin, host-specific variation, stability over time, and as yet, no consistent patterns differentiating psoriatic skin from healthy skin.Human skin is colonized by diverse microbiota, including bacteria and fungi, that can be pathogenic under particular circumstances (14, 16). Traditionally, microorganisms have been identified by culture-dependent methods; however, many species are fastidious and underrepresented in cultures from mixed microbial communities (13), whereas others cannot be cultivated under known conditions (2). Therefore, culture-independent molecular techniques have been used for the identification of microbial species within ecosystems (2, 9, 27, 42).Such methods, particularly the analysis of rRNA genes, have been employed to characterize bacterial and fungal communities associated with diverse human body sites, including intestine (11), gingiva (28,33,43), esophagus (45), vagina (65), and outer ear canal (13). As predicted, these studies revealed greater diversity, including previously undescribed organisms, than did previous analyses based on culture-dependent techniques.The application of molecular techniques has been advocated to characterize the microbiota in both healthy and diseased skin (14). To date, rRNA data have been used to identify species associated with fungal dermatoses (21,29,38,39) and PCR-based diagnostic tests have been developed (15,26,62). Psoriasis, a common dermatosis affecting about 3% of the population in industrialized countries (3), is characterized by erythrosquamous cutaneous lesions associated with abnormal patterns of keratinocyte growth and differentiation (35). Although of unknown etiology, trigger factors, including physical trauma and streptococcal infections, may provoke clinical manifestations (51). Fungal organisms, including Candida albicans (63) and Malassezia furfur (3), have also been associated with the developmen...

show abstract

“…The Western blot technique using antigen components from 22-70 kDa is able to detect antibodies to major proteins at 40-70 kDa in patients with sporotrichosis [23] and also differentiate between cutaneous and extracutaneous forms by showing darker bands for extracutaneous sporotrichosis, as compared to the cutaneous type [23]. Molecular methods such as nested polymerase chain reaction [25] and restriction fragment length polymorphism (RFLP) analysis [26,27] are no doubt very sensitive and instrumental in diagnosis of disease when the organism is in a very small amount such as cerebrospinal fluid (CSF). But due to their high costs and the skilled expertise required to perform them, they will remain primarily research tools for epidemiological investigations.…”

Section: Discussionmentioning

confidence: 99%

A case series and review of sporotrichosis in Sikkim

Bhutia¹,

Gurung²,

Yegneswaran

et al. 2011

J Infect Dev Ctries

View full text Add to dashboard Cite

Sporotrichosis caused by the fungus Sporothrix schenckii has been widely reported from the northern Himalayan belt and the north eastern region of India. Three autochthonous cases of lymphocutaneous sporotrichosis from east and south districts of Sikkim are reported. Fluid aspirate from the nodulo-ulcerative lesions were sent for cytology and fungal culture. S. schenckii was isolated on culture and cytological examination in all three cases showed granulomatous reaction. Thermal dimorphism was demonstrated and animal pathogenicity testing was performed. Saturated solution of potassium iodide was used for treatment and the last case was treated with itraconazole and potassium iodide. Awareness of this disease and an extensive environmental study is required to understand the actual burden of this disease.

show abstract

Detection of Sporothrix schenckii in Clinical Samples by a Nested PCR Assay

Cited by 74 publications

References 23 publications

Rapid Identification of Sporothrix Species by T3B Fingerprinting

Rapid Identification of Sporothrix Species by T3B Fingerprinting

Molecular Analysis of Fungal Microbiota in Samples from Healthy Human Skin and Psoriatic Lesions

A case series and review of sporotrichosis in Sikkim

Contact Info

Product

Resources

About