The principles of idiotypic (Id) vaccination were used to immunize against vaginitis caused by Ca... more The principles of idiotypic (Id) vaccination were used to immunize against vaginitis caused by Candida albicans, a widespread and sometimes intractable disease in women. To this aim, a murine mAb (KT4, IgG1) neutralizing in vitro the anti-Candida activity of a yeast killer toxin (YKT) was used as an Id vaccine to elicit Abs with toxin-like activity in a rat vaginitis model. Nonimmunized and isotype-matched, irrelevant mAb-immunized rats served as controls. An effective protection was obtained in Id-vaccinated animals, as demonstrated by a highly significant decrease in vaginal Candida CFU compared with controls. The protection was associated with rising vaginal titers of anti-idiotypic Abs (IdAb), prevalently of the IgA isotype, that were able to passively transfer the protective state to nonimmunized animals. The vaginal IdAb possessed YKT-like activity because they were able to kill in vitro the challenging fungal cells, and this killing was neutralized by the mAb KT4. Overall, th...
A single primer pair amplifying a cytochrome P-450 lanosterol-14 alpha-demethylase (L1A1) gene fr... more A single primer pair amplifying a cytochrome P-450 lanosterol-14 alpha-demethylase (L1A1) gene fragment that encodes a highly conserved region was used to detect yeast DNA in clinical specimens. Positive PCR products were obtained from genomic DNAs of Candida albicans, C. parapsilosis, C. tropicalis, C. guilliermondii, C. krusei, C. (Torulopsis) glabrata, and C. kefyr. No human, bacterial, or parasitic DNA was amplified. The sensitivity was evaluated for C. albicans genomic DNA by using various DNA concentrations (200 pg to 2 fg). The amplified DNAs of Candida species with unknown P-450 L1A1 gene sequences were subcloned and sequenced. Identification at the species level was achieved by digestion of the PCR products with different restriction enzymes. A specific restriction enzyme analysis pattern was determined for each species investigated. Subsequently, we used PCR to detect specific yeast DNA directly with clinical specimens such as blood and bronchoalveolar lavage specimens. Af...
Blood samples were drawn daily from 72 patients who had hematological malignancies, neutropenia, ... more Blood samples were drawn daily from 72 patients who had hematological malignancies, neutropenia, and fever and who had failed to respond to broad-spectrum antibiotics. Each sample was used for conventional fungal blood cultures and for detection and identification of Candida DNA by a PCR method with subsequent restriction enzyme analysis (REA) recently developed in our laboratory. The PCR method was able to detect five CFU of Candida spp. per ml of blood, and subsequent REA of the amplicons allowed the identification of the Candida species most commonly implicated in cases of candidiasis. Thirty-one patients were PCR-REA positive, and four of these patients were also culture positive. The ultimate diagnosis for 13 of these patients and 1 patient who was PCR-REA negative was disseminated candidiasis (confirmed by clinical data, multiple cultures, histology, autopsy, and/or ultrasonographic evidence of hepatosplenic candidiasis). The molecular method is significantly more sensitive th...
A PCR-based assay was developed to detect and identify medically important yeasts in clinical sam... more A PCR-based assay was developed to detect and identify medically important yeasts in clinical samples. Using a previously described set of primers (G. Morace et al., J. Clin. Microbiol. 35:667–672, 1997), we amplified a fragment of the ERG11 gene for cytochrome P-450 lanosterol 14α-demethylase, a crucial enzyme in the biosynthesis of ergosterol. The PCR product was analyzed in a reverse cross blot hybridization assay with species-specific probes directed to a target region of the ERG11 gene of Candida albicans (pCal), C. guilliermondii (pGui), C. ( Torulopsis ) glabrata (pGla), C. kefyr (pKef), C. krusei (pKru), C. parapsilosis (pPar), C. tropicalis (pTro), the newly described species C. dubliniensis (pDub), Saccharomyces cerevisiae (pSce), and Cryptococcus neoformans (pCry). The PCR-reverse cross blot hybridization assay correctly identified multiple isolates of each species tested. No cross-hybridization was detected with any other fungal, bacteria, or human DNAs tested. The metho...
The differential action of four selected yeast killer toxins on the mycelial and yeast forms of f... more The differential action of four selected yeast killer toxins on the mycelial and yeast forms of four isolates of the dimorphic fungus Sporothrix schenckii was comparatively evaluated. The results confirmed that the yeast killer phenomenon is present among hyphomycetes and yeasts and that both morphological forms of S. schenckii are susceptible to the action of the same yeast killer toxin. Quantitative differences in the response to the killer action of the mycelial and yeast forms in individual strains were also observed. To avoid retroconversion of the dimorphic forms, we used a modification of the conventional killer system.
Cultures of Petriellidium boydii were serologically identified by detection of their exoantigens ... more Cultures of Petriellidium boydii were serologically identified by detection of their exoantigens with an immunodiffusion procedure. The technique, which is specific and sensitive, allowed the rapid identification and differentiation of 12 isolates of P. boydii from numerous other morphologically similar Hyphomycetes. The antigen-antiserum reference system and the production, by two different techniques, of exoantigens used in the identification of P. boydii are described.
Summary. Candida spp. (83 isolates including C. (Torulopsis) glabrata) were tested in vitro for t... more Summary. Candida spp. (83 isolates including C. (Torulopsis) glabrata) were tested in vitro for their susceptibility to 5‐fluorocytosine, amphotericin B, ketoconazole, itraconazole, fluconazole, and miconazole. The yeasts were isolated from clinical specimens, mostly from the lower respiratory tract, of 30 oncologic patients, 27/30 with haematological malignancies, during a 6‐month period (December 1991‐May 1992). Minimal inhibitory concentration (MIC) and minimal fungicidal concentration (MFC) values of the 6 drugs were obtained for each yeast using a microdilution broth method developed in our laboratory. Amphotericin B, and 5‐fluorocytosine were active against the majority of the yeasts with MIC90/MFC90 values within achievable serum concentrations (3.12/6.25 μg ml‐1 and 0.625/0.625 μg ml‐1 respectively). Azole derivatives showed a species‐specific activity. MFC values were two to four times higher than those of the MICs, confirming the fungistatic rather than fungicidal activity...
Summary: Trichophyton mentagrophytes and Microsporum canis were isolated from apparently healthy ... more Summary: Trichophyton mentagrophytes and Microsporum canis were isolated from apparently healthy scalps of schoolboys during an epidemiological study of dermatophyte carriers. Various other keratinophilic fungi were also isolated.Zusammenfassung: Während einer epidemiologischen Untersuchung auf Dermatophyten‐Träger konnten Trichophyton mentagrophytes and Microsporum canis von offensichtlich gesunder Kopfhaut bei Schuljungen isoliert werden.
A Rhodococcus equi pulmonary infection in a 63-year-old man receiving chemotherapy and radiothera... more A Rhodococcus equi pulmonary infection in a 63-year-old man receiving chemotherapy and radiotherapy for spinocellular carcinoma is described. The patient, a knife-grinder, was promptly treated with levofloxacin plus amikacin followed by rifampicin for 2 months, and he is still in good clinical condition after an 8-month follow-up.
Fluconazole susceptibility among 800 clinical Candida isolates (60% C. albicans ) and two control... more Fluconazole susceptibility among 800 clinical Candida isolates (60% C. albicans ) and two control strains ( C. krusei ATCC 6258 and C. parapsilosis ATCC 22019) was tested with the NCCLS M27-A method (gold standard) and six commercial products (Candifast, disk, Etest, Fungitest, Integral System Yeasts, and Sensititre YeastOne). Results were classified as susceptible, susceptible-dose dependent, or resistant using M27-A breakpoints or, for Fungitest, Integral System Yeasts, and Candifast, as susceptible, intermediate, or resistant, according to the manufacturers' instructions. Concordance with NCCLS M27-A results was analyzed with the χ 2 test. Intra- and interlaboratory reproducibility was also evaluated. NCCLS M27-A (90.1%), Etest (93.1%), Sensititre YeastOne (93.1%), disk (96.7%), Fungitest (92.6%), Integral System Yeasts (40.6%), and Candifast (6.0%) classified the indicated percentages of C. albicans isolates as susceptible. Among non- C. albicans strains, the percentages of ...
Infection Control & Hospital Epidemiology, 1990
Eight patients in two surgical units developed systemic candidosis during a 40-day period from Ju... more Eight patients in two surgical units developed systemic candidosis during a 40-day period from June 5 to July 13, 1987 (in five casesCandida albicanswas identified). Three of them died. All cases belonged to a group of 27 patients receiving parenteral nutrition (PN), while among the 108 patients who did not receive PN, no cases were observed (p = .000001).Candidawas cultured from two PN bags administered to the cases. A specialized nutrition nurse was responsible for the PN compounding and for maintaining administration sets in the two wards involved.An epidemiological investigation in which 19 uninfected patients who had had PN were used as controls, showed no significant difference between cases and controls except that lipids were more frequently added to bags administered to cases (p = .0005). Furthermore, the bags administered to cases contained a higher average number of multidose constituents (p = .0008) when the comparison was focused on the two days before the onset of symp...
The principles of idiotypic (Id) vaccination were used to immunize against vaginitis caused by Ca... more The principles of idiotypic (Id) vaccination were used to immunize against vaginitis caused by Candida albicans, a widespread and sometimes intractable disease in women. To this aim, a murine mAb (KT4, IgG1) neutralizing in vitro the anti-Candida activity of a yeast killer toxin (YKT) was used as an Id vaccine to elicit Abs with toxin-like activity in a rat vaginitis model. Nonimmunized and isotype-matched, irrelevant mAb-immunized rats served as controls. An effective protection was obtained in Id-vaccinated animals, as demonstrated by a highly significant decrease in vaginal Candida CFU compared with controls. The protection was associated with rising vaginal titers of anti-idiotypic Abs (IdAb), prevalently of the IgA isotype, that were able to passively transfer the protective state to nonimmunized animals. The vaginal IdAb possessed YKT-like activity because they were able to kill in vitro the challenging fungal cells, and this killing was neutralized by the mAb KT4. Overall, th...
A single primer pair amplifying a cytochrome P-450 lanosterol-14 alpha-demethylase (L1A1) gene fr... more A single primer pair amplifying a cytochrome P-450 lanosterol-14 alpha-demethylase (L1A1) gene fragment that encodes a highly conserved region was used to detect yeast DNA in clinical specimens. Positive PCR products were obtained from genomic DNAs of Candida albicans, C. parapsilosis, C. tropicalis, C. guilliermondii, C. krusei, C. (Torulopsis) glabrata, and C. kefyr. No human, bacterial, or parasitic DNA was amplified. The sensitivity was evaluated for C. albicans genomic DNA by using various DNA concentrations (200 pg to 2 fg). The amplified DNAs of Candida species with unknown P-450 L1A1 gene sequences were subcloned and sequenced. Identification at the species level was achieved by digestion of the PCR products with different restriction enzymes. A specific restriction enzyme analysis pattern was determined for each species investigated. Subsequently, we used PCR to detect specific yeast DNA directly with clinical specimens such as blood and bronchoalveolar lavage specimens. Af...
Blood samples were drawn daily from 72 patients who had hematological malignancies, neutropenia, ... more Blood samples were drawn daily from 72 patients who had hematological malignancies, neutropenia, and fever and who had failed to respond to broad-spectrum antibiotics. Each sample was used for conventional fungal blood cultures and for detection and identification of Candida DNA by a PCR method with subsequent restriction enzyme analysis (REA) recently developed in our laboratory. The PCR method was able to detect five CFU of Candida spp. per ml of blood, and subsequent REA of the amplicons allowed the identification of the Candida species most commonly implicated in cases of candidiasis. Thirty-one patients were PCR-REA positive, and four of these patients were also culture positive. The ultimate diagnosis for 13 of these patients and 1 patient who was PCR-REA negative was disseminated candidiasis (confirmed by clinical data, multiple cultures, histology, autopsy, and/or ultrasonographic evidence of hepatosplenic candidiasis). The molecular method is significantly more sensitive th...
A PCR-based assay was developed to detect and identify medically important yeasts in clinical sam... more A PCR-based assay was developed to detect and identify medically important yeasts in clinical samples. Using a previously described set of primers (G. Morace et al., J. Clin. Microbiol. 35:667–672, 1997), we amplified a fragment of the ERG11 gene for cytochrome P-450 lanosterol 14α-demethylase, a crucial enzyme in the biosynthesis of ergosterol. The PCR product was analyzed in a reverse cross blot hybridization assay with species-specific probes directed to a target region of the ERG11 gene of Candida albicans (pCal), C. guilliermondii (pGui), C. ( Torulopsis ) glabrata (pGla), C. kefyr (pKef), C. krusei (pKru), C. parapsilosis (pPar), C. tropicalis (pTro), the newly described species C. dubliniensis (pDub), Saccharomyces cerevisiae (pSce), and Cryptococcus neoformans (pCry). The PCR-reverse cross blot hybridization assay correctly identified multiple isolates of each species tested. No cross-hybridization was detected with any other fungal, bacteria, or human DNAs tested. The metho...
The differential action of four selected yeast killer toxins on the mycelial and yeast forms of f... more The differential action of four selected yeast killer toxins on the mycelial and yeast forms of four isolates of the dimorphic fungus Sporothrix schenckii was comparatively evaluated. The results confirmed that the yeast killer phenomenon is present among hyphomycetes and yeasts and that both morphological forms of S. schenckii are susceptible to the action of the same yeast killer toxin. Quantitative differences in the response to the killer action of the mycelial and yeast forms in individual strains were also observed. To avoid retroconversion of the dimorphic forms, we used a modification of the conventional killer system.
Cultures of Petriellidium boydii were serologically identified by detection of their exoantigens ... more Cultures of Petriellidium boydii were serologically identified by detection of their exoantigens with an immunodiffusion procedure. The technique, which is specific and sensitive, allowed the rapid identification and differentiation of 12 isolates of P. boydii from numerous other morphologically similar Hyphomycetes. The antigen-antiserum reference system and the production, by two different techniques, of exoantigens used in the identification of P. boydii are described.
Summary. Candida spp. (83 isolates including C. (Torulopsis) glabrata) were tested in vitro for t... more Summary. Candida spp. (83 isolates including C. (Torulopsis) glabrata) were tested in vitro for their susceptibility to 5‐fluorocytosine, amphotericin B, ketoconazole, itraconazole, fluconazole, and miconazole. The yeasts were isolated from clinical specimens, mostly from the lower respiratory tract, of 30 oncologic patients, 27/30 with haematological malignancies, during a 6‐month period (December 1991‐May 1992). Minimal inhibitory concentration (MIC) and minimal fungicidal concentration (MFC) values of the 6 drugs were obtained for each yeast using a microdilution broth method developed in our laboratory. Amphotericin B, and 5‐fluorocytosine were active against the majority of the yeasts with MIC90/MFC90 values within achievable serum concentrations (3.12/6.25 μg ml‐1 and 0.625/0.625 μg ml‐1 respectively). Azole derivatives showed a species‐specific activity. MFC values were two to four times higher than those of the MICs, confirming the fungistatic rather than fungicidal activity...
Summary: Trichophyton mentagrophytes and Microsporum canis were isolated from apparently healthy ... more Summary: Trichophyton mentagrophytes and Microsporum canis were isolated from apparently healthy scalps of schoolboys during an epidemiological study of dermatophyte carriers. Various other keratinophilic fungi were also isolated.Zusammenfassung: Während einer epidemiologischen Untersuchung auf Dermatophyten‐Träger konnten Trichophyton mentagrophytes and Microsporum canis von offensichtlich gesunder Kopfhaut bei Schuljungen isoliert werden.
A Rhodococcus equi pulmonary infection in a 63-year-old man receiving chemotherapy and radiothera... more A Rhodococcus equi pulmonary infection in a 63-year-old man receiving chemotherapy and radiotherapy for spinocellular carcinoma is described. The patient, a knife-grinder, was promptly treated with levofloxacin plus amikacin followed by rifampicin for 2 months, and he is still in good clinical condition after an 8-month follow-up.
Fluconazole susceptibility among 800 clinical Candida isolates (60% C. albicans ) and two control... more Fluconazole susceptibility among 800 clinical Candida isolates (60% C. albicans ) and two control strains ( C. krusei ATCC 6258 and C. parapsilosis ATCC 22019) was tested with the NCCLS M27-A method (gold standard) and six commercial products (Candifast, disk, Etest, Fungitest, Integral System Yeasts, and Sensititre YeastOne). Results were classified as susceptible, susceptible-dose dependent, or resistant using M27-A breakpoints or, for Fungitest, Integral System Yeasts, and Candifast, as susceptible, intermediate, or resistant, according to the manufacturers' instructions. Concordance with NCCLS M27-A results was analyzed with the χ 2 test. Intra- and interlaboratory reproducibility was also evaluated. NCCLS M27-A (90.1%), Etest (93.1%), Sensititre YeastOne (93.1%), disk (96.7%), Fungitest (92.6%), Integral System Yeasts (40.6%), and Candifast (6.0%) classified the indicated percentages of C. albicans isolates as susceptible. Among non- C. albicans strains, the percentages of ...
Infection Control & Hospital Epidemiology, 1990
Eight patients in two surgical units developed systemic candidosis during a 40-day period from Ju... more Eight patients in two surgical units developed systemic candidosis during a 40-day period from June 5 to July 13, 1987 (in five casesCandida albicanswas identified). Three of them died. All cases belonged to a group of 27 patients receiving parenteral nutrition (PN), while among the 108 patients who did not receive PN, no cases were observed (p = .000001).Candidawas cultured from two PN bags administered to the cases. A specialized nutrition nurse was responsible for the PN compounding and for maintaining administration sets in the two wards involved.An epidemiological investigation in which 19 uninfected patients who had had PN were used as controls, showed no significant difference between cases and controls except that lipids were more frequently added to bags administered to cases (p = .0005). Furthermore, the bags administered to cases contained a higher average number of multidose constituents (p = .0008) when the comparison was focused on the two days before the onset of symp...
Uploads
Papers by G. Morace