scholarly journals A Reference Laboratory Surveillance on Fungal Isolates from Patients with Haematological Malignancy in Japan

2021 ◽  
Vol 7 (10) ◽  
pp. 806
Author(s):  
Yutaro Hino ◽  
Akira Watanabe ◽  
Rio Seki ◽  
Shokichi Tsukamoto ◽  
Yusuke Takeda ◽  
...  
Keyword(s):  
Fusarium Species ◽  
Invasive Fungal Disease ◽  
Underlying Disease ◽  
Drug Susceptibility ◽  
Rapid Identification ◽  
Haematopoietic Stem Cell ◽  
Causative Organism ◽  
Reference Laboratory ◽  
Laboratory Surveillance ◽  
Causative Fungus

Invasive fungal disease (IFD) in patients with haematological disorders is a fatal disease, making rapid identification and treatment crucial. However, the identification of the causative fungus is often difficult, sometimes even impossible. There have been few reports concerning the causative species of IFD. This study aimed to investigate the epidemiology and causative organism of IFD in patients with haematological diseases in Japan. We analyzed the IFD cases among the patients with haematological malignancies identified at the Medical Mycological Research Center, Chiba University, between 2013 and 2019. The most common underlying disease was acute myeloid leukaemia (34.3%). Forty-six point one percent of IFD patients received haematopoietic stem cell transplantation (HSCT). The major pathogens were Aspergillus, Candida, and Fusarium. Aspergillus fumigatus was the most common Aspergillus species, and Candida fermentati and Fusarium petroliphilum were the most common Candida and Fusarium species, respectively, in this analysis. Furthermore, various cryptic species and non-albicans Candida were identified. The drug susceptibility of such relatively rare strains suggests that analysis of the causative fungi should provide valuable information for therapeutic options. Therefore, our study indicated that it is clinically significant to identify the organism in as much detail as possible.

scholarly journals Type A7 Coxsackie (type 4 poliomyelitis) virus infection in Scotland

1962 ◽  
Vol 60 (3) ◽  
pp. 323-332 ◽  
Author(s):  
N. R. Grist
Keyword(s):  
Tissue Culture ◽  
Blood Transfusion ◽  
Virus Infection ◽  
Technical Assistance ◽  
Animal Experiments ◽  
Rapid Identification ◽  
National Institutes Of Health ◽  
Reference Laboratory ◽  
Blood Transfusion Service ◽  
The Many

Coxsackie A 7 virus was isolated from thirty-seven patients during an outbreak in Scotland in 1959. Seven cases were paralytic, one of them fatal. Evidence is presented that Coxsackie A 7 virus caused these paralytic illnesses. The virus was also isolated from a paralytic case in 1956 and from a non-paralytic case in 1961. Serological surveys suggest that it has been active in the community for some years. Specific haemagglutination by Coxsackie A 7 virus was useful for rapid identification of viruses and for measurement of serum antibodies.I am grateful to Dr A. D. Macrae of the Virus Reference Laboratory, Colindale, London, for prototype Coxsackie A 7 virus; to Dr K. Habel of the National Institutes of Health, Bethesda, U.S.A., for tissue culture-adapted ABIV virus; to Dr J. Wallace of the Blood Transfusion Service, West of Scotland Region, for samples of blood donor sera; to Dr M. Rentsch, Klinik für Kinderkrankheiten, University of Berne, Switzerland, for permission to quote the results of virological tests of his cases; to Miss R. McLelland, F.A.T.A., and to Mr C. McLean F.I.M.L.T., for technical assistance with animal experiments; to Mr H. G. Carson, F.I.M.L.T. and to Mr J. Kerr, A.I.M.L.T., for technical assistance with neutralization tests; and to the many clinical colleagues who provided specimens and information for this study.

LAMP assay for distinguishing Fusarium oxysporum and Fusarium commune in lotus (Nelumbo nucifera) rhizomes

Plant Disease ◽  
2021 ◽  
Author(s):  
Sheng Deng ◽  
Xin Ma ◽  
Yifan Chen ◽  
Hui Feng ◽  
Dongmei Zhou ◽  
...  
Keyword(s):  
Fusarium Oxysporum ◽  
Fusarium Species ◽  
Lamp Assay ◽  
Rapid Identification ◽  
Nelumbo Nucifera ◽  
Disease Spread ◽  
Molecular Diagnostic ◽  
Primer Sets ◽  
Rot Disease ◽  
Rhizome Rot

The yields of edible rhizome from the cultivation of the perennial hydrophyte lotus (Nelumbo nucifera) can be severely reduced by rhizome rot disease caused by Fusarium species. There is a lack of rapid field-applicable methods for detection of these pathogens on lotus plants displaying symptoms of rhizome-rot. Fusarium commune (91%) and Fusarium oxysporum (9%) were identified at different frequencies from lotus samples showing symptoms of rhizome-rot. As these two species can cause different severity of disease and their morphology is very similar, molecular-diagnostic based methods to detect these two species were developed. Based on the comparison of the mitochondrial genome of the two species, three specific DNA loci targets were found. The designed primer sets for conventional PCR, qPCR and loop-mediated isothermal amplification (LAMP) precisely distinguished the above two species when isolated from lotus and other plants. The LAMP detection limits were 10 pg/μl and 1 pg/μl of total DNA for F. commune and F. oxysporum, respectively. We also carried out field-mimicked experiments on lotus seedlings and rhizomes (including inoculated samples and field diseased samples), and the results indicated that the LAMP primer sets and the supporting portable methods are suitable for the rapid diagnosis of the lotus disease in the field. The LAMP-based detection method will aid in the rapid identification of whether F. oxysporum or F. commune are infecting lotus plants with symptoms of rhizome-rot, and can facilitate efficient pesticide use and prevent the disease spread through vegetative propagation of Fusarium-infected lotus rhizomes.

scholarly journals Invasive Haemophilus influenzae disease in adults

2000 ◽  
Vol 124 (3) ◽  
pp. 441-447 ◽  
Author(s):  
J. SARANGI ◽  
K. CARTWRIGHT ◽  
J. STUART ◽  
S. BROOKES ◽  
R. MORRIS ◽  
...  
Keyword(s):  
Haemophilus Influenzae ◽  
Clinical Presentation ◽  
Age Groups ◽  
Case Fatality ◽  
Underlying Disease ◽  
Reference Laboratory ◽  
Age Group ◽  
Childhood Immunization ◽  
Clinical Presentations ◽  
Vaccines For Children

We reviewed retrospectively all invasive Haemophilus influenzae (Hi) infections in adults ascertained from reference laboratory records and notifications from five NHS regions over the 5 years from 1 October 1990, a period encompassing the introduction of routine Hib childhood immunization (October 1992). A total of 446 cases were identified, a rate of 0·73 infections per 105 adults per annum. Though numbers of Hib infections in adults fell after the introduction of Hib vaccines for children (P = 0·035), and there was no increase in infections caused by other capsulated Hi serotypes, total numbers of invasive Hi infections increased due to a large rise in infections caused by non-capsulated Hi (ncHi) strains (P = 0·0067). There was an unexpectedly low rate of infections in those aged 75 years or more (P < 0·0001). The commonest clinical presentations were pneumonia with bacteraemia (227/350, 65%) and bacteraemia alone (62/350, 18%) and the highest rates of disease were in the 65–74 years age group (P < 0·0001). Clinical presentation was not influenced by the capsulation status of the invading Hi strain. 103/350 cases (29%) died within 1 month, and 207/350 (59%) within 6 months of their Hi infection. Case fatality rates were high in all age groups. Pre-existing diseases were noted in 220/350 cases and were associated with a higher case fatality rate (82% vs. 21%, P < 0·0001). After the introduction of Hib immunization in children, invasive Hib infections in unimmunized adults also declined, but the overall rate of invasive Hi disease in adults increased, with most infections now caused by non-capsulated strains. Physicians and microbiologists should be aware of the changing epidemiology, the high associated mortality and high risk of underlying disease. Invasive haemophilus infections in adults should be investigated and treated aggressively.

scholarly journals Real time PCR for the rapid identification and drug susceptibility of Mycobacteria present in Bronchial washings

2016 ◽  
Vol 16 (1) ◽  
Author(s):  
Thilini Piushani Keerthirathne ◽  
Dhammika Nayoma Magana-Arachchi ◽  
Dushantha Madegedara ◽  
Suneth Sithumini Sooriyapathirana
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Drug Susceptibility ◽  
Rapid Identification

Proficiency testing of conventional drug susceptibility tests of Mycobacterium tuberculosis

1987 ◽  
Vol 33 (12) ◽  
pp. 1064-1068 ◽  
Author(s):  
Adalbert Laszlo ◽  
Dorothy M. Helbecque ◽  
Walter Tostowaryk
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Proficiency Testing ◽  
Susceptibility Testing ◽  
Developed Countries ◽  
Drug Susceptibility ◽  
Reference Laboratory ◽  
In Vitro Susceptibility ◽  
Testing Procedures ◽  
Drug Concentrations ◽  
Susceptibility Tests

Proficiency testing of indirect drug susceptibility tests of Mycobacterium tuberculosis was begun in 1985 by the Laboratory Centre for Disease Control (LCDC) with the participation of Provincial Public Health Laboratories in Canada. Comparable sets of 60 cultures of Mycobacterium tuberculosis representing 30 strains were distributed by LCDC to the participating laboratories to be tested for drug susceptibility against isoniazid, streptomycin, rifampin, and ethambutol using conventional methodologies. Intralaboratory agreement values determined by comparing results obtained on sets of duplicate cultures were high and were found to vary little from drug to drug and from laboratory to laboratory. Interlaboratory agreement was determined by comparing results reported by participating laboratories to those obtained by the Reference Laboratory. Agreement percentages were found to be lower for drug-resistant cultures than for drug-susceptible cultures. The reliability of drug susceptibility testing results was higher for isoniazid and rifampin, than for ethambutol and streptomycin. This study shows that the higher subsidiary drug concentrations do not compare well with main drug concentrations, especially in the case of streptomycin and ethambutol. The significance of the higher subsidiary concentrations in in vitro susceptibility testing is therefore in need of clarification. The proficiency testing results obtained in this study compare favorably with those reported in other developed countries despite the fact that a variety of testing procedures are used throughout the country.

scholarly journals Speeding up the diagnosis of multidrug-resistant tuberculosis in a high-burden region with the use of a commercial line probe assay

2019 ◽  
Vol 45 (2) ◽  
Author(s):  
Angela Pires Brandao ◽  
Juliana Maira Watanabe Pinhata ◽  
Rosangela Siqueira Oliveira ◽  
Vera Maria Neder Galesi ◽  
Helio Hehl Caiaffa-Filho ◽  
...  
Keyword(s):  
Susceptibility Testing ◽  
Drug Susceptibility ◽  
Multidrug Resistant ◽  
Drug Susceptibility Testing ◽  
Reference Laboratory ◽  
Line Probe Assay ◽  
Resistant Tuberculosis ◽  
Multidrug Resistant Tuberculosis ◽  
Commercial Line ◽  
Probe Assay

ABSTRACT Objective: To evaluate the rapid diagnosis of multidrug-resistant tuberculosis, by using a commercial line probe assay for rifampicin and isoniazid detection (LPA-plus), in the routine workflow of a tuberculosis reference laboratory. Methods: The LPA-plus was prospectively evaluated on 341 isolates concurrently submitted to the automated liquid drug susceptibility testing system. Results: Among 303 phenotypically valid results, none was genotypically rifampicin false-susceptible (13/13; 100% sensitivity). Two rifampicin-susceptible isolates harboured rpoB mutations (288/290; 99.3% specificity) which, however, were non-resistance-conferring mutations. LPA-plus missed three isoniazid-resistant isolates (23/26; 88.5% sensitivity) and detected all isoniazid-susceptible isolates (277/277; 100% specificity). Among the 38 (11%) invalid phenotypic results, LPA-plus identified 31 rifampicin- and isoniazid-susceptible isolates, one isoniazid-resistant and six as non-Mycobacterium tuberculosis complex. Conclusions: LPA-plus showed excellent agreement (≥91%) and accuracy (≥99%). Implementing LPA-plus in our setting can speed up the diagnosis of multidrug-resistant tuberculosis, yield a significantly higher number of valid results than phenotypic drug susceptibility testing and provide further information on the drug-resistance level.

scholarly journals GenoType NTM-DR Performance Evaluation for Identification of Mycobacterium avium Complex and Mycobacterium abscessus and Determination of Clarithromycin and Amikacin Resistance

2019 ◽  
Vol 57 (8) ◽  
Author(s):  
Hee Jae Huh ◽  
Su-Young Kim ◽  
Hyang Jin Shim ◽  
Dae Hun Kim ◽  
In Young Yoo ◽  
...  
Keyword(s):  
Mycobacterium Avium ◽  
Drug Susceptibility ◽  
Rapid Identification ◽  
Drug Susceptibility Testing ◽  
Mycobacterium Abscessus ◽  
Clinical Isolates ◽  
Resistance Mutations ◽  
Content Type ◽  
Clarithromycin Resistance ◽  
Reference Strains

ABSTRACT We evaluated the GenoType NTM-DR (NTM-DR) line probe assay for identifying Mycobacterium avium complex (MAC) species and Mycobacterium abscessus subspecies and for determining clarithromycin and amikacin resistance. Thirty-eight reference strains and 145 clinical isolates (58 MAC and 87 M. abscessus isolates), including 54 clarithromycin- and/or amikacin-resistant strains, were involved. The performance of the NTM-DR assay in rapid identification was evaluated by comparison with results of multigene sequence-based typing, whereas performance in rapid detection of clarithromycin and amikacin resistance was evaluated by comparison with sequencing of the erm(41), rrl, and rrs genes and drug susceptibility testing (DST). The accuracies of MAC and M. abscessus (sub)species identification were 92.1% (35/38) and 100% (145/145) for the 38 reference strains and 145 clinical isolates, respectively. Three MAC strains other than M. intracellulare were found to cross-react with the M. intracellulare probe in the assay. Regarding clarithromycin resistance, NTM-DR detected rrl mutations in 52 isolates and yielded 99.3% (144/145) and 98.6% (143/145) concordant results with sequencing and DST, respectively. NTM-DR sensitivity and specificity in the detection of clarithromycin resistance were 96.3% (52/54) and 100% (91/91), respectively. The NTM-DR yielded accurate erm(41) genotype results for all 87 M. abscessus isolates. Regarding amikacin resistance, NTM-DR detected rrs mutations in five isolates and yielded 99.3% (144/145) and 97.9% (142/145) concordant results with sequencing and DST, respectively. Our results indicate that the NTM-DR assay is a straightforward and accurate approach for discriminating MAC and M. abscessus (sub)species and for detecting clarithromycin and amikacin resistance mutations and that it is a useful tool in the clinical setting.

scholarly journals Rapid Identification of Fungi by Using the ITS2 Genetic Region and an Automated Fluorescent Capillary Electrophoresis System

1999 ◽  
Vol 37 (6) ◽  
pp. 1846-1851 ◽  
Author(s):  
Christine Y. Turenne ◽  
Steven E. Sanche ◽  
Daryl J. Hoban ◽  
James A. Karlowsky ◽  
Amin M. Kabani
Keyword(s):  
Fungal Infections ◽  
Invasive Fungal Disease ◽  
Rapid Identification ◽  
Molecular Techniques ◽  
Its2 Region ◽  
Promising Tool ◽  
Fungal Isolates ◽  
Identification Of Fungi ◽  
Clinically Significant ◽  
Species Specific

Invasive fungal disease often plays an important role in the morbidity and mortality of immunocompromised patients. The poor sensitivity of current fungal blood culture and histological practices has led to the development of highly sensitive and specific molecular techniques, such as the PCR. Sequence variability of the internal transcribed spacer 2 (ITS2) region of fungi is potentially useful in rapid and accurate diagnosis of clinical fungal isolates. PCR with fungus-specific primers targeted toward conserved sequences of the 5.8S and 28S ribosomal DNA (rDNA) results in amplification of the species-specific ITS2 regions, which are variable in amplicon length. We have made use of the ABI PRISM 310 genetic analyzer and the ABI PRISM 310 GeneScan analysis software for the determination of variable size differences of the ITS2 region of clinically important fungi, including Candida and non-Candida yeasts,Aspergillus species, and a variety of dermatophytes. No cross-reaction occurred when samples were tested against human and bacterial genomic DNA. We have found that most clinically significant fungal isolates can be differentiated by this method, and it therefore serves to be a promising tool for the rapid (<7 h) diagnosis of fungemia and other invasive fungal infections.

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