scholarly journals An alarming rise of non-albicans Candida species and uncommon yeasts in the clinical samples; a combination of various molecular techniques for identification of etiologic agents

2019 ◽  
Author(s):  
Mohireh Taei ◽  
Mostafa Chadeganipour ◽  
Rasoul Mohammadi
Keyword(s):  
Candida Species ◽  
Systemic Infection ◽  
Molecular Techniques ◽  
Clinical Isolates ◽  
Clinical Samples ◽  
Molecular Tests ◽  
Yeast Strains ◽  
Etiologic Agents ◽  
Pcr Multiplex ◽  
Pcr Rflp

Abstract Objective: Yeasts are unicellular microorganisms may cause systemic infection in immunocompromised patients. The aim of this study was to identify yeast strains isolated from clinical specimens using molecular techniques. Results: A total of 202 yeast strains isolated from 341 clinical samples between February 2017 and May 2019. All clinical isolates were identified using phenotypic and molecular tests including PCR-RFLP, duplex-PCR, multiplex-PCR, and PCR-sequencing. The most yeast fungal isolates were obtained from urine (66.8%), nail (9.4%), skin lesion (7.9%), bronchoalveolar lavage (5.9%), and blood (3.9%). One hundred and twenty-one Candida species were identified as non- albicans versus 76 Candida albicans. Trichosporon asahii, and Pichia terricola were uncommon non- Candida yeasts isolated from urine samples. For the first time, we isolated P. terricola as etiological agent of urinary tract infection in a pregnant female. Since Candida species show different levels of resistance to antifungal agents, precise identification of clinical isolates is critical for better treatment of infection.

scholarly journals An alarming rise of non-albicans Candida species and uncommon yeasts in the clinical samples; a combination of various molecular techniques for identification of etiologic agents

2019 ◽  
Author(s):  
Mohireh Taei ◽  
Mostafa Chadeganipour ◽  
Rasoul Mohammadi
Keyword(s):  
Candida Species ◽  
Systemic Infection ◽  
Molecular Techniques ◽  
Clinical Isolates ◽  
Clinical Samples ◽  
Molecular Tests ◽  
Yeast Strains ◽  
Etiologic Agents ◽  
Pcr Multiplex ◽  
Pcr Rflp

Abstract Objective: Yeasts are unicellular microorganisms may cause systemic infection in immunocompromised patients. The aim of this study was to identify yeast strains isolated from clinical specimens using molecular techniques. Results: A total of 202 yeast strains isolated from 341 clinical samples between February 2017 and May 2019. All clinical isolates were identified using phenotypic and molecular tests including PCR-RFLP, duplex-PCR, multiplex-PCR, and PCR-sequencing. The most yeast fungal isolates were obtained from urine (66.8%), nail (9.4%), skin lesion (7.9%), bronchoalveolar lavage (5.9%), and blood (3.9%). One hundred and twenty-one Candida species were identified as non-albicans versus 76 Candida albicans. Trichosporon asahii, and Pichia terricola were uncommon non-Candida yeasts isolated from urine samples. For the first time, we isolated P. terricola as etiological agent of urinary tract infection in a pregnant female. Since Candida species show different levels of resistance to antifungal agents, precise identification of clinical isolates is critical for better treatment of infection.

scholarly journals An alarming rise of non-albicans Candida species and uncommon yeasts in the clinical samples; a combination of various molecular techniques for identification of etiologic agents

2019 ◽  
Author(s):  
Mohireh Taei ◽  
Mostafa Chadeganipour ◽  
Rasoul Mohammadi
Keyword(s):  
Candida Species ◽  
Systemic Infection ◽  
Molecular Techniques ◽  
Clinical Isolates ◽  
Clinical Samples ◽  
Molecular Tests ◽  
Yeast Strains ◽  
Etiologic Agents ◽  
Pcr Multiplex ◽  
Pcr Rflp

Abstract Objective: Yeasts are unicellular microorganisms may cause systemic infection in immunocompromised patients. The aim of this study was to identify yeast strains isolated from clinical specimens using molecular techniques. Results: A total of 202 yeast strains isolated from 341 clinical samples between February 2017 and May 2019. All clinical isolates were identified using phenotypic and molecular tests including PCR-RFLP, duplex-PCR, multiplex-PCR, and PCR-sequencing. The most yeast fungal isolates were obtained from urine (66.8%), nail (9.4%), skin lesion (7.9%), bronchoalveolar lavage (5.9%), and blood (3.9%). One hundred and twenty-one Candida species were identified as non- albicans versus 76 Candida albicans. Trichosporon asahii, and Pichia terricola were uncommon non- Candida yeasts isolated from urine samples. For the first time, we isolated P. terricola as etiological agent of urinary tract infection in a pregnant female. Since Candida species show different levels of resistance to antifungal agents, precise identification of clinical isolates is critical for better treatment of infection.

scholarly journals An alarming rise of non-albicans Candida species and uncommon yeasts in the clinical samples; a combination of various molecular techniques for identification of etiologic agents

2019 ◽  
Vol 12 (1) ◽  
Author(s):  
Monireh Taei ◽  
Mostafa Chadeganipour ◽  
Rasoul Mohammadi
Keyword(s):  
Systemic Infection ◽  
Molecular Techniques ◽  
Clinical Isolates ◽  
Clinical Samples ◽  
Molecular Tests ◽  
Yeast Strains ◽  
Etiologic Agents ◽  
Pcr Multiplex ◽  
Pcr Rflp ◽  
First Time

Abstract Objective Yeasts are unicellular microorganisms may cause systemic infection in immunocompromised patients. The aim of this study was to identify yeast strains isolated from clinical specimens using molecular techniques. Results A total of 202 yeast strains isolated from 341 clinical samples between February 2017 and May 2019. All clinical isolates were identified using phenotypic and molecular tests including PCR–RFLP, duplex-PCR, multiplex-PCR, and PCR-sequencing. The most yeast fungal isolates were obtained from urine (66.8%), nail (9.4%), skin lesion (7.9%), bronchoalveolar lavage (5.9%), and blood (3.9%). One hundred and twenty-one Candida species were identified as non-albicans versus 76 Candida albicans. Trichosporon asahii, and Pichia terricola were uncommon non-Candida yeasts isolated from urine samples. For the first time, we isolated P. terricola as etiological agent of urinary tract infection in a pregnant female. Since Candida species show different levels of resistance to antifungal agents, precise identification of clinical isolates is critical for better treatment of infection.

scholarly journals An alarming rise of non-albicans Candida species and uncommon yeasts in the clinical samples; a combination of various molecular techniques for identification of etiologic agents

2019 ◽  
Author(s):  
Mohireh Taei ◽  
Mostafa Chadeganipour ◽  
Rasoul Mohammadi
Keyword(s):  
Molecular Identification ◽  
Candida Species ◽  
Fungal Infections ◽  
Antifungal Susceptibility ◽  
Etiologic Agent ◽  
Molecular Techniques ◽  
University Hospital ◽  
Medical Mycology ◽  
Clinical Samples ◽  
Pcr Multiplex

Abstract Objective: Yeasts are opportunistic microorganisms can cause human fungal infection among immunocompromised patients. This study aimed to identify Candida species and uncommon yeasts obtained from clinical specimens in Kashani university hospital and Shefa Lab as a referral medical mycology laboratory, in Isfahan, Iran, by combination of various molecular techniques. Results: A total of 202 yeast strains were isolated from 341 clinical samples between February 2017 to May 2019. All clinical isolates were identified using phenotypic and molecular tests. PCR-RFLP, duplex-PCR, multiplex-PCR, and PCR-sequencing were applied for molecular identification of yeasts. The most clinical samples were obtained from urine (66.8%), nail (9.4%), bronchoalveolar lavage (5.9%), sore (4.4%), and blood (3.9%). One hundred and twenty-one Candida species were identified as non- albicans against 76 Candida albicans. Trichosporon asahii, and Pichia terricola were uncommon non- Candida yeasts isolated from urine samples. For the first time, we isolated P. terricola as etiologic agent of urinary tract infection in a pregnant female. Since non- albicans Candida species and non- Candida yeasts have various virulence factors and antifungal susceptibility profile, precise molecular identification can help us to reach to the advantageous strategies for treatment of these fungal infections.

scholarly journals Identification of Candida Species from Clinical Samples in a Honduran Tertiary Hospital

2019 ◽  
Author(s):  
Kathy Montes ◽  
Bryan Ortiz ◽  
Celeste Galindo ◽  
Isis Figueroa ◽  
Sharleen Braham ◽  
...  
Keyword(s):  
Candida Species ◽  
Dna Amplification ◽  
Tertiary Hospital ◽  
Clinical Samples ◽  
Identification Methods ◽  
Pcr Rflp ◽  
Fungal Dna ◽  
Genotypic Identification ◽  
Human Infections ◽  
And Control

Candida species are one of the most important causes of human infections, especially in hospitals and among immunocompromised patients. The correct and rapid etiological identification of yeast infections is important to provide adequate therapy, reduce mortality and control outbreaks. In this study, Candida species were identified in patients with suspected fungal infection, and phenotypic and genotypic identification methods were compared. A total of 167 axenic fungal cultures and 46 clinical samples were analyzed by HardyCHROM®, MicroScan®, and PCR-RFLP. The species of the C. albicans complex were the most frequent, followed by C. tropicalis and C. glabrata. Less common but clinically relevant species of Candida were also isolated. The comparison between the three methods was concordant, especially for the most common Candida species. Fungal DNA amplification was successful in all clinical samples.

Molecular identification of some Hoplolaimus species from the USA based on duplex PCR, multiplex PCR and PCR-RFLP analysis

Nematology ◽  
2009 ◽  
Vol 11 (3) ◽  
pp. 471-480 ◽  
Author(s):  
Robert Robbins ◽  
Allen Szalanski ◽  
Chang-Hwan Bae
Keyword(s):  
Multiplex Pcr ◽  
Dna Sequences ◽  
Rflp Analysis ◽  
Pcr Primers ◽  
Interspecific Variation ◽  
Molecular Techniques ◽  
Specific Pcr ◽  
Pcr Multiplex ◽  
Pcr Rflp ◽  
Species Specific

AbstractTwo different molecular approaches, a multiplex PCR and PCR-RFLP of ITS-rDNA, were developed for the identification of Hoplolaimus species. DNA sequences of H. columbus, H. galeatus, H. concaudajuvencus, H. magnistylus, H. seinhorsti and three undescribed Hoplolaimus species were used to design species-specific primers. Three reverse species-specific PCR primers for H. columbus, H. galeatus and H. magnistylus were developed using the ITS1 region exhibiting interspecific variation. Three species-specific PCR primers in combination with the forward primer, Hoc-1f, produced distinct amplicons of 580 bp for H. columbus, 120 bp for H. galeatus and 340 bp for H. magnistylus. We successfully identified each of three species by multiplex PCR when all three were mixed in a single PCR reaction. Restriction enzyme digests of the PCR amplicon using HaeIII and RsaI permitted discrimination of H. columbus, H. galeatus, H. magnistylus, H. concaudajuvencus, H. sp. 1, H. sp. 2 and H. sp. 3 from each other. These results suggest that these molecular techniques allow for rapid, easy and reliable identification of Hoplolaimus species.

scholarly journals Distribution of pathogenicity islands among Colombian isolates of Salmonella

2010 ◽  
Vol 4 (09) ◽  
pp. 555-559 ◽  
Author(s):  
Miryan Margot Sánchez-Jiménez ◽  
Nora María Cardona-Castro ◽  
Nunzia Canu ◽  
Sergio Uzzau ◽  
Salvatore Rubino
Keyword(s):  
Systemic Infection ◽  
Clinical Isolates ◽  
Clinical Samples ◽  
Pathogenicity Islands ◽  
Gastrointestinal Infection ◽  
Heterogeneous Distribution ◽  
Geographical Regions ◽  
Stool Samples ◽  
Salmonella Pathogenicity Islands ◽  
Reference Strains

Introduction: Salmonella pathogenicity islands (SPIs) are regions scattered along the bacterial chromosome, with an acknowledged pivotal role during gastrointestinal and systemic infection. The distribution of SPIs has been investigated in reference strains. However, there is a lack of studies on their presence and/or assortment within the genomes of Salmonella enterica (S. enterica) serovars that circulate in different geographical regions. Therefore, in this study, we aimed to determine the presence of genes of the pathogenicity islands 1 to 5 (SPI-1 to 5), in Salmonella clinical isolates from Colombian patients with systemic and enteric outcomes. Methodology: A total of 125 strains of S. enterica belonging to different serovars were isolated from various clinical samples. Strains were identified and screened for the presence of various genes located in pathogenicity islands. The genes tested were selected according to the attributed pathogenic function and detected by PCR for the SPI-1 hilA and invA; for SPI-2 spiC and ttrC; for SPI-3 misL and mgtC; for SPI-4 orfL and SPI-4R; and for SPI-5 pipD and sopB. Results: Salmonella pathogenicity islands 1 to 5 were detected in isolates from patients with systemic and gastrointestinal infection. All the systemic isolates possessed all the genes tested; in contrast, 16 isolates from stool samples lacked one or more sequences encoded by the SPI-3 and SPI-4 (p < 0.000001). Conclusions: These results describe the heterogeneous distribution of SPIs-encoded sequences within the genomes of Colombian clinical isolates, and reveal important differences among systemic and stool sample isolates.

scholarly journals Identification of Candida Species from Clinical Samples in a Honduran Tertiary Hospital

Pathogens ◽  
2019 ◽  
Vol 8 (4) ◽  
pp. 237 ◽  
Author(s):  
Kathy Montes ◽  
Bryan Ortiz ◽  
Celeste Galindo ◽  
Isis Figueroa ◽  
Sharleen Braham ◽  
...  
Keyword(s):  
Candida Species ◽  
Dna Amplification ◽  
Tertiary Hospital ◽  
Clinical Samples ◽  
Length Polymorphisms ◽  
Pcr Rflp ◽  
Fungal Dna ◽  
Genotypic Identification ◽  
Human Infections ◽  
And Control

Candida species are one of the most important causes of human infections, especially in hospitals and among immunocompromised patients. The correct and rapid etiological identification of yeast infections is important to provide adequate therapy, reduce mortality, and control outbreaks. In this study, Candida species were identified in patients with suspected fungal infection, and phenotypic and genotypic identification methods were compared. A total of 167 axenic fungal cultures and 46 clinical samples were analyzed by HardyCHROM®, MicroScan®(Omron Microscan Systems Inc, Renton, WA, USA), and PCR-RFLP (Restriction Fragment Length Polymorphisms). The species of the C. albicans complex were the most frequent, followed by C. tropicalis and C. glabrata. Less common but clinically relevant species of Candida were also isolated. The comparison between the three methods was concordant, especially for the most common Candida species. Fungal DNA amplification was successful in all clinical samples.

scholarly journals A STUDY ON IDENTIFICATION AND SPECIATION OF MEDICALLY IMPORTANT CANDIDA SPECIES ISOLATED FROM VARIOUS CLINICAL SAMPLES BY USING HICROME CANDIDA.

2020 ◽  
pp. 1-3
Author(s):  
Mr Jogender ◽  
Anjali Kulshrestha* ◽  
Suman Rishi
Keyword(s):  
Candida Albicans ◽  
Conventional Method ◽  
Candida Species ◽  
Antifungal Agents ◽  
Clinical Isolates ◽  
Clinical Samples ◽  
Clinical Specimens ◽  
Major Species ◽  
Conventional Methods ◽  
Isolated Species

Candida albicans remains the most frequently isolated species, but an increase in the prevalence of Non-albicans candida is a matter of concern for 1-3 various laboratories, as NAC show less susceptibility to antifungal agents particularly azoles . Routinely used conventional methods are cumbersome and time consuming. Hence the present study aimed at species identication of candida isolated from various clinical specimens and also to evaluate the usefulness of HiCrome Candida differential agar as compared to routine conventional method for speciation of candida. Atotal of 51 non repetitive clinical isolates of Candida were obtained from various clinical samples. Candida albicans was the major species accounting for 23 (45.10%) of the total isolates. Non albicans Candida constituted 17 (33.33%) of C. tropicalis, 9 (17.64%) of C. krusei and 2 (3.92%) of C. parasilosis. 49 Candida species which were correctly identied by HiCrome Candida agar except 2 species of C.parasilosis (identied by conventional method) which were identied as C. glabrata by Hicrome Candida agar

scholarly journals Development and validation of a multiplex qPCR assay for detection and relative quantification of HPV16 and HPV18 E6 and E7 oncogenes

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Alexia Bordigoni ◽  
Anne Motte ◽  
Hervé Tissot-Dupont ◽  
Philippe Colson ◽  
Christelle Desnues
Keyword(s):  
Limit Of Detection ◽  
Quantitative Polymerase Chain Reaction ◽  
Human Papillomaviruses ◽  
Molecular Techniques ◽  
Clinical Samples ◽  
Gene Encoding ◽  
Hpv Dna ◽  
E6 And E7 ◽  
High Risk Hpv ◽  
Development And Validation

AbstractHuman papillomaviruses (HPV) play a key role in promoting human anogenital cancers. Current high-risk HPV screening or diagnosis tests involve cytological or molecular techniques mostly based on qualitative HPV DNA detection. Here, we describe the development of a rapid quantitative polymerase chain reaction (qPCR) detection test of HPV16 and HPV18 oncogenes (E6 and E7) normalized on human gene encoding GAPDH. Optimized qPCR parameters were defined, and analytical specificities were validated. The limit of detection was 101 for all genes tested. Assay performances were evaluated on clinical samples (n = 96). Concordance between the Xpert HPV assay and the triplex assay developed here was 93.44% for HPV16 and 73.58% for HPV18. HPV co-infections were detected in 15 samples. The systems developed in the present study can be used in complement to traditional HPV tests for specifically validating the presence of HPV16 and/or HPV18. It can also be used for the follow-up of patients with confirmed infection and at risk of developing lesions, through the quantification of E6 and E7 oncogene expression (mRNA) normalized on the GAPDH expression levels.

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