Nanthawan Mekha

Evaluation of a Newly Developed Lateral Flow Immunoassay for the Diagnosis of Cryptococcosis

This study, evaluating the performance of a novel cryptococcal lateral flow immunoassay, shows that the assay performs as well as available diagnostic methods is economical, rapid, and easy to perform; and as such can be a point of care test in resource limited settings.

The First Isolation of Ustilaginomycetous Anamorphic Yeasts, Pseudozyma Species, from Patients' Blood and a Description of Two New Species: P. parantarctica and P. thailandica

The genus Pseudozyma is ustilaginomycetous anamorphic yeasts, and are mainly isolated from plants. We isolated three Pseudozyma strains from the blood of patients in Thailand. While one isolate was identified as P. antarctica by rDNA sequence analysis, the other two were considered to be new species and were named P. parantarctica and P. thailandica. The three isolates proved to be resistant to 5‐flucytosine, and P. thailandica was also resistant to fluconazole and itraconazole. As far as we know, this is the first isolation of Pseudozyma strains from humans. The two new species are described.

Serotyping of Cryptococcus neoformans strains isolated from clinical specimens in Thailand and their susceptibility to various antifungal agents

One hundred and thirty-nine strains of Cryptococcus neoformans were isolated in Thailand from clinical specimens including 97 AIDS patients: 67 from Northern, 48 from Central, 17 from Northeastern and 7 from Southern regional hospitals. Six out of the 139 strains were serotype B and the remaining 133 were A. There was no correlation between serotypes and regional distribution. To our knowledge, this is the first report of serotyping studies on C. neoformans in Thailand. Studies on random amplified polymorphic DNA (RAPD) analysis showed that this method is useful for the differentiation of C. neoformans var. gattii (serotypes B and C) and C. neoformans var. neoformans (serotypes A and D). They also indicated that Thai isolates of C. neoformans var. gattii (serotype B) were a homogeneous group on the basis of their genotypes. Antifungal susceptibility tests using 5 antifungal agents including amphotericin B, fluconazole, flucytosine, itraconazole and micronazole against 50 selected strains of C. neoformans showed that they were sensitive to all of the antifungal agents tested except for one strain that was resistant to flucytosine.

Characterization of clinical isolates of pathogenic Nocardia strains and related actinomycetes in Thailand from 1996 to 2003

In Thailand from 1996 to 2003, 171 strains of pathogenic aerobic actinomycetes from clinical specimens were isolated. Of those strains, 134 were mycolic acid containing actinomycetes, including 96 strains of Nocardia species. Others included 10 strains of Gordonia, 14 strains of Rhodococcus, and 22 strains of Mycobacterium. One strain each of the genera Tsukamurella and Corynebacterium were also isolated. Also identified were 27 strains of non-mycolic acid containing actinomycetes. Our identification studies of 96 strains of Nocardia species showed that significant pathogens in Thailand were N. beijingensis (18 strains), N. cyriacigeorgica (13 strains), and N. farcinica (34 strains); the most prevalent species was N. farcinica (35.4%). We also isolated four strains of N. asiatica, five strains of N. asteroides sensu stricto, four strains of N. nova, seven strains of N. otitidiscaviarum, eight strains of N. transvalensis, and two strains of N. pseudobrasiliensis.

Candida pseudohaemulonii Sp. Nov., an Amphotericin B‐ and Azole‐Resistant Yeast Species, Isolated from the Blood of a Patient from Thailand

Candida haemulonii (types I and II) is rarely isolated from clinical specimens. We isolated a strain that is phylogenetically close to C. haemulonii from the blood of a Thai patient, and named it C. pseudohaemulonii sp. nov. (CBS 10099T=JCM 12453T=DMST 17134T). The new species and C. haemulonii types I and II were resistant to amphotericin B and azole agents but were susceptible to a 1,3‐β‐D‐glucan synthetase inhibitor, micafungin, and 5‐flucytosine. The species were easily distinguished using an ID32 yeast identification kit. The taxonomic description of C. pseudohaemulonii sp. nov. is presented.

Genotyping and antifungal drug susceptibility of the pathogenic yeast Trichosporon asahii isolated from Thai patients.

Trichosporonosis due to Trichosporon asahii is a life-threatening infection with a very poor prognosis. We analyzed the genotype of intergenic transcribed spacer (IGS) region 1 of the rRNA gene and determined the drug susceptibility of 101 T. asahii isolates obtained from Thai patients to collect basic information on trichosporonosis in Thailand. Of the five genotypes in the IGS region identified in this study, types 1 and 3 were predominant in Thailand. The distribution in Thailand differs from that in other countries, suggesting that there is a geographic substructure among T. asahii clinical isolates. Voriconazole appeared to be the most active drug.

In vitro antifungal activity of the new triazole D0 870 against Penicillium marneffei compared with that of amphotericin B, fluconazole, itraconazole, miconazole and flucytosine

Summary. The in vitro activity of D0870, a new triazole, was compared with that of fluconazole, kraconazole, miconazole, amphotericin B and flucytosine against recent clinical isolates of Penicillium marneffei in Thailand. The minimum inhibitory concentrations (MIC) values were determined by a microbroth dilution method using morpholinopropanesulphonic acid (MOPS)‐buffered RPMI‐1640 and brain heart infusion (BHI) medium. Yeast nitrogen base (YNB) medium with glucose was also used for the assay of flucytosine. D0870 was less active against P. marneffei than itraconazole, but its activity was similar to that of miconazole, superior to that of amphotericin B and markedly superior to that of fluconazole and flucytosine. Much lower MIC values of D0870 were observed with BHI medium.

Three new basidiomycetous yeasts, Pseudozyma alboarmeniaca sp. nov., Pseudozyma crassa sp. nov. and Pseudozyma siamensis sp. nov. isolated from Thai patients.

We previously reported the first isolation of Pseudozyma species from the blood of Thai patients. In this study, three additional new Pseudozyma species were isolated from clinical specimens from Thai patients. The Pseudozyma species showed relatively low sensitivity to azole antifungal agents. The names proposed for these isolates are Pseudozyma alboarmeniaca (DMST 17135T = JCM 12454T = CBS 9961T), Pseudozyma crassa (DMST 17136T = JCM 12455T = CBS 9959T) and Pseudozyma siamensis (DMST 17137T = JCM 12456T CBS 9960T), where DMST is Department of Medical Sciences Culture Collection, JCM is Japan Collection of Microorganisms and CBS is Centraalbureau voor Schimmelcultures.

Random amplified polymorphic DNA (RAPD) analysis of Penicillium marneffei strains isolated from AIDS patients in Thailand

Results of random amplified polymorphic DNA (RAPD) analysis using three different primers showed that 16 strains ofPenicillium marneffei isolated from AIDS patients in Thailand belonged to a genetically homogenous group, but different slightly from an isolate from bamboo rat in China. Six PCR fragments (from about, 200 to 600 bp) that were commonly observed in the RAPD fingerprint of all strains were extracted and sequented. Usefulness of this sequence information for identification ofP. marneffei is discussed.

Comparative evaluation of Trichosporon asahii susceptibility using ASTY colorimetric microdilution and CLSI M27-A2 broth microdilution reference methods

The in vitro activity of AMPH‐B, 5‐FC, FLCZ, MCZ, ITCZ, and VRCZ against 50 isolates of T. asahii was determined using CLSI M27‐A2 microdilution and ASTY colorimetric methods. Observed agreement ranged from 96 to 100% according to the drug. Overall, the agreement between two methods was 97.7%. The ASTY colorimetric method was thus determined to be comparable to the CLSI reference method when testing the susceptibility of T. asahii to a variety of antifungal agents.