共查询到20条相似文献,搜索用时 31 毫秒
1.
Penicilliosis marneffei, often consecutive to the aspiration of Talaromyces marneffei (Penicillium marneffei), continues to be one of the significant causes of morbidity and mortality in immunocompromised patients in endemic regions such as Southeast Asia. Improving the accuracy of diagnosing this disease would aid in reducing the mortality of associated infections. In this study, we developed a stable and reproducible murine pulmonary model that mimics human penicilliosis marneffei using a nebulizer to deliver Talaromyces marneffei (SUMS0152) conidia to the lungs of BALB/c nude mice housed in exposure chamber. Using this model, we further revealed that nested PCR was sensitive and specific for detecting Talaromyces marneffei in bronchoalveolar lavage fluid and fresh tissues. This inhalation model may provide a more representative analysis tool for studying the development of penicilliosis marneffei, in addition to revealing that nested PCR has a predictive value in reflecting pulmonary infection. 相似文献
2.
Deborah A. Cortlandt José Luis López-Ribot K. John Morrow David C. Straus W. LaJean Chaffin 《Mycopathologia》1995,131(1):1-8
Penicilliosis marneffei has emerged as an endemic systemic mycosis in Southeast Asia among humans and wild bamboo rats. To gain an insight into the epidemiology of this life-threatening disease, a survey of bamboo rats for natural infections byPenicillium marneffei was carried out in the central plains of Thailand during June-September, 1987. Thirty-one lesser bamboo rats (Cannomys badius) and eight hoary bamboo rats (Rhizomys pruinosus) were trapped. Portions of their internal organs were cultured to determine if they had been infected byP. marneffei. Six each ofC. badius (19.4%) andR. pruinosus (75%) yielded cultures of this unique, dimorphicPenicillium species. All of the isolates were readily converted to their unicellular form that multiplies by the process of schizogony by incubating them at 37 °C on plates of brain heart infusion agar. Their identity was further confirmed by a specific immunological test. Among the internal organs of the positive rats, the lungs had the highest positivity (83.3%), next in decreased order of frequency were the liver (33.3%) and the pancreas (33.3%). The use and value of domestic and wild animals in locating and demarcating endemic areas of geophilic fungal pathogens are discussed. Penicilliosis marneffei is considered to be a zooanthroponosis — a disease that occurs in lower animals, as well as, humans. 相似文献
3.
Penicillium marneffei: types and drug susceptibility 总被引:6,自引:0,他引:6
Imwidthaya P Thipsuvan K Chaiprasert A Danchaivijitra S Sutthent R Jearanaisilavong J 《Mycopathologia》2001,149(3):109-115
The PCR fingerprints of 30 Penicillium marneffei isolates from Chiang Rai in northern Thailand and Bangkok in central Thailand were studied through use of single-nucleotide
primers (GACA)4 and the phage M13 core sequence. Discrimination of fingerprint patterns was based on differences in the number of major bands.
The P. marneffei isolates were divided into four types, i.e., A, B, C, and D. Type A was found in two isolates from Chiang Rai (6.7%). Types
B and C respectively were found in two (6.7%) and one (3.3%) isolates from Bangkok. The predominate type D (83.3%) was found
in isolates obtained from Chiang Rai and Bangkok. The PCR fingerprinting method was found to be useful for the epidemiological
study of P. marneffei, a dimorphic opportunistic fungus and an emerging pathogen in the HIV pandemic. In vitro drug susceptibility testing by broth macrodilution to four antifungal agents against the yeast form ofP. marneffei was performed. The MIC ranges for amphotericin B, fluconazole, itraconazole, and ketoconazole were 0.125–0.5, 4.0–8.0, <0.032,
and <0.125 μg/ml respectively.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
4.
该研究采用稀释涂布法结合形态观察、16S rRNA基因序列分析,对广西北海川蔓藻(Ruppia maritima)内生及根际细菌的物种多样性进行了研究,并采用琼脂扩散法和光度计法分析了其粗提物抑制马尔尼菲青霉菌活性。结果表明:从川蔓藻中分离到可培养内生细菌26株,根际可培养细菌31株。分别将内生细菌归属为10科12属13种,根际分离出细菌归属为9科14属19种,其中5株根际细菌可能为潜在新种。获得8株细菌对马尔尼菲青霉菌有抑制活性,总阳性率为25.0%。其中,菌株BGMRC 2015、BGMRC 2059、BGMRC 2043的粗提物表现出较强的抑制马尔尼菲青霉菌效果,其MIC分别为(1.800±0.045)、(1.881±0.061)、(1.604±0.021)mg·m L~(-1)。川蔓藻中可培养细菌具有较高的物种多样性,蕴藏着丰富的新物种资源,且富含抑菌活性良好的菌株。 相似文献
5.
Molecular methods were carried out to detect Penicillium griseofulvum, a dominant species related to heavy metal pollution, which was screened from marine contaminated sediments. Based on differences
in internal transcribed spacer (ITS) sequences of Penicillium genus and specific isoamyl alcohol oxidase (IAO) sequences, species-specific primers AS1/RS4 and IAO1/IAO2 of Penicillium griseofulvum were designed and synthesized which were then employed in optimized PCR systems. The detection sensitivities were compared
through ordinary PCR and nested-PCR using two pairs of primers, respectively. Both primer pairs could exclusively amplify
destined DNA fragment from contaminated environmental samples in our researches. As for primers AS1/RS4, the detection sensitivity
for spores (pure spore DNA) could be 10 fg/μl and 10 spores, respectively, and the detection sensitivity for the sediments
was 102 spores/0.25 g sediments. While the detection sensitivity of IAO1/IAO2 primers was lower than that of AS1/RS4. Despite the
difference in detection sensitivity, it is feasible that the species-specific primers could be used as probes for the detection
of environmental pollution dominant species, Penicillium griseofulvum, since the frequency of occurrence and amount of this strain could preferably indicate the pollution degree. 相似文献
6.
Infection by Penicillium marneffei is an important emerging public health problem, especially among travelers and inhabitants in SE China and SE Asia infected
with human immunodeficiency virus (HIV). In recent years, the number of patients with penicilliosis marneffei (PM) has increased
rapidly in Guangdong province, SE China. However, the natural habitat and transmission mode of the etiologic agent remains
unclear. In this study, wild rats (Microtus, focus Rattus and Rhizomys pruinosus) and soil samples were collected from rat
burrows, populated and rural areas from November 2007 to December 2008 for fungus cultures. All isolates, suspected of being
P. marneffei, were identified by gross and microscopic morphology and ITS analysis. Sixteen of 23 (about 70%) bamboo rats were P. marneffei positive, whereas none was recovered from hamsters, loirs or soil. This suggests that as of today the bamboo rat is the exclusive
natural reservoir for P. marneffei. Definite evidence that rodents are a part of the infectious cycle is still lacking. 相似文献
7.
Nanthawan Mekha Natteewan Poonwan Dr. Yuzuru Mikami Katsukiyo Yazawa Tohru Gonoi Shuji Hasegawa Kazuko Nishimura 《Mycoscience》1997,38(2):97-100
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. 相似文献
8.
Cunwei Cao Glenn Bulmer Jushang Li Ling Liang Youkun Lin Yongjia Xu Qinghua Luo 《Mycopathologia》2010,170(1):47-50
This is the first indigenous case of disseminated histoplasmosis reported from the Penicillium marneffei endemic area in southern China. It was diagnosed by histopathology of tissue, gross and microscopic morphology of the culture
and PCR assay of the isolated fungus. Successful antifungal treatment was with itraconazole 400 mg/day for 5 months. This
case suggests that histoplasmosis should be an important differential diagnosis in immunocompromised patients in southern
China and South East Asia (the only endemic area for P. marneffei). 相似文献
9.
10.
Ying-ge Wang Jin-mei Cheng Hai-bo Ding Xi Lin Guo-hao Chen Mei Zhou Sheng-nan Ye 《Mycopathologia》2018,183(3):551-558
Objective
To improve the diagnosis and treatment of Penicilliosis marneffei without human immunodeficiency virus infection.Methods
Analyze and review the clinical features, diagnosis and treatment of six cases of P. marneffei without human immunodeficiency virus infection at The First Affiliated Hospital of Fujian Medical University.Results
Two cases were diagnosed in the ENT Department, three cases in the respiratory department and one case in the dermatological department. Penicillium marneffei infection was confirmed by sputum culture, blood culture and tissue biopsy. After definite diagnosis, one refused further treatment, and others showed significant improvement.Conclusion
Penicilliosis marneffei is insidious onset and easy to be escaped and misdiagnosed. To achieve early diagnosis and appropriate treatment, doubtful cases should be alerted for the diagnoses as P. marneffei.11.
An immunodiffusion (ID) test has been developed to diagnose infections caused byPenicillium marneffei. A 20 X concentrated culture-filtrate of six-week-old shake cultures (25 C) ofP. marneffei was employed as an antigen. This preparation was found to be better in quality than that from still cultures of the same age (30 C). Anti-P. marneffei rabbit sera were produced by injecting rabbits with increasing dosages of the inoculum for at least six weeks. These sera demonstrated two to three precipitin lines following their reaction with their antigens for 24–48 h at 25 C. TheP. marneffei antigenic preparations did not react with rabbit antisera to five species ofAspergillus, which commonly cause aspergillosis, or to antisera for four dimorphic systemic fungi. Similarly, the antigens of these other fungi did not react against the anti-P. marneffei rabbit serum. However, the anti-P. marneffei rabbit sera demonstrated antibody titres (132 to 164) to histoplasmin, blastomycin and coccidioidin in the complement fixation test. Cross-reactions were not observed with any of the human sera in the suspected or proven cases of opportunistic or systemic mycotic infections. Therefore, the ID test for penicillosis marneffei is considered to be highly specific. Exoantigen studies demonstrated that, of a total of 34 isolates of ten species ofPenicillium tested, only the extracts ofP. marneffei (6 isolates) and one isolate (PLM 771) of aP. species reacted positively with the anti-P. marneffei rabbit serum, giving at least two lines of identity with reference reagents. Based on this analysis, theP. species (PLM 771) was identified as P. marneffei. The exoantigen test is considered to be a specific and rapid method for the identification and confirmation ofP. marneffei isolates.
Portion of a thesis submitted by the second author to the Canadian Society of Laboratory Technologists, Hamilton, Ontario, Canada, in partial fulfilment of the requirements for Advanced registered Technologist, Certification in Immunology. 相似文献
Zusammenfassung Eine Immun-Verbreitung (ID) Test war ausgearbeitet fur die Diagnose der Infektionen diePenicillium marneffei verursachen. Die 20 X conzentrierte Kultur-Fieltrier von der sechs Wochen alten schuttelten Kulturen (25 C) vonP. Marneffei war gebraucht fur Antigen. Diese Preparation war besser in Qualitat denn die stillen Kulturen von dem selben Alter (30 C). Die Anti-P. Marneffei Kaninchen Sera wurde produziert durch die Injektion der Kaninchen mit immer vergrosserden Dosen von dem Imfstoff-mindestebs wahrend sechs Wochen. Diese Sera zeigte zwei — drei Ubere sturzung Linien durch die Reaktion von ihrer Antigenen wahrend 24–48 Stunden im 25 C. Die Antigen-Preparationen vonP. Marneffei gaben keine Reaktionen mit Kaninchen Anti-sera fur funf Species vonAspergillus, die stellen an gewohnlich Aspergillosis, oder zu der Antisera fur fier dimorpische systemische Pilze. Gleichweis, die Antigenen von diese Plize gaben keine Reaktionen gegen die Kaninchen Antisera vonP. Marneffei. Jedoch die Kaninchen Antiserum vonP. marneffei zeigte Anti-Korper Titer (132 bis 164) zu Histoplasmin, Blastomycin und Coccidioidin in der Complement (Erganzung) Fixing Test. Kreuz-Reaktionen wurden nicht observiert mit keine menschlichen Sera in verdachtigen oder erwiesenen Krankengeschichten von passenden oder systematischen mycotischen Infectionen. Deswegen die ID Test fur Penicillosis marneffei ist sehr spezifisch. Die Untersuchungen haben demonstriert, dass 34 isolationen von zehn Species vonPenicillium — die untersucht waren — nur der Auszug vonP. Marneffei (6 Isolationen) und ein Isolation (PLM 771) vonP. Species eine positive Reaktion mit der Kaninchen Anti-Serum vonP. Marneffei, und es gab mindestens zwei identische Linen mit die Referenz Reagenten. An diese Analyse grunden wir dass, dieP. Species (PLM 771) war identifiziert wieP. Marneffei. Die Exo-Antigen Test ist anbetrachtet wie eine spezifische und schnelle Methode fur die Identifikation und Bestatigung der Isolierten vonP. Marneffei.
Portion of a thesis submitted by the second author to the Canadian Society of Laboratory Technologists, Hamilton, Ontario, Canada, in partial fulfilment of the requirements for Advanced registered Technologist, Certification in Immunology. 相似文献
12.
Penicilliosis is a disease caused by Penicillium marneffei, a fungus endemic to Southeast Asia. Prior to the HIV/AIDS epidemic, infection was exceedingly rare, but penicilliosis is
currently one of the most common opportunistic infections in persons with HIV/AIDS in some Asian countries. This paper describes
the clinical manifestations, diagnosis, and epidemiology of this emerging opportunistic infection and will focus on some gaps
in our knowledge and directions for future research. 相似文献
13.
Zhang JM Sun JF Feng PY Li XQ Lu CM Lu S Cai WY Xi LY de Hoog GS 《Journal of microbiological methods》2011,85(1):33-39
Penicillium marneffei infection is a deadly disease and early diagnosis leads to prompt and appropriate antifungal therapy. To develop a sensitive method to diagnose P. marneffei infection, a multiplex ligation-dependent probe amplification (MLPA) assay was adapted. This method can rapidly and specifically detect P. marneffei DNA in cultured cells and paraffin-embedded tissue samples. Three pairs of probes were designed for amplifying the internally (intergenic) transcribed spacer (ITS) region of P. marneffei rRNA using a systematic phylogenetic analysis. These three probe sets produced three amplicons of 198, 166, and 152 bp, respectively, specific for P. marneffei. In contrast, there was only one 198 bp amplicon produced for Talaromyces stipitatus, and one 152 bp amplicon for P. funiculosum, T. intermedius and T. derxii. The probes did not amplify any other reference strains. An array of 40 P. marneffei strains isolated from human patients, bamboo rat, and the local environment was tested by using MLPA, and all were positively identified. Most importantly, P. marneffei in paraffin-embedded tissue specimens from infected human patients was positively amplified by MLPA. The sensitivity and specificity of the MLPA assay could be a useful tool for prompt diagnosis, pathogen characterization, and epidemiological studies of fungal infections. 相似文献
14.
The ITS region of ectomycorrhizal fungi was analyzed, and species-specific PCR primers were designed for 8 ectomycorrhizal
Tricholoma species. Although a high degree of intraspecific homology was observed, interspecific variation was sufficient to design
species-specific primers based on sequence of the ITS region. PCR amplification with the specific primers generated fragments
of the expected sizes from DNA extracted from the strains of each species but gave no amplified products from the strains
of the other 16 species in eight genera. These results suggest that sequence of the ITS region is appropriate to be used for
species-level identification of ectomycorrhizal fungi. 相似文献
15.
16.
Detection of hydA genes of Clostridia spp. using degenerative and species specific primers for C. butyricum were optimized by the addition of bovine serum albumin (BSA) to polymerase chain reaction (PCR) and quantitative PCR (qPCR)
reactions. BSA concentrations ranging from 100 to 400 ng/μl were examined using pure cultures and a variety of environmental
samples as test targets. A BSA concentration of 100 ng/μl, which is lower than previously reported in the literature, was
found to be most effective in improving the detection limit. The brightness of amplicons with 100 ng/μl BSA increased in ethidium
bromide-treated gels, the minimum detection limit with BSA was at least one log greater, and cycle threshold (C
T) values were lower than without BSA in qPCR indicating improved detection of target deoxyribonucleic acid for most samples
tested. Although amplicon visualization was improved at BSA concentrations greater than or equal to 100 ng/μl, gene copy numbers
detected by qPCR were less, CT values were increased, and T
m values were altered. SYBR Green dissociation curves of qPCR products of DNA from pure culture or sludge samples showed that
BSA at 100 ng/μl reduced the variability of peak areas and T
m values. 相似文献
17.
We have developed a polymerase chain reaction (PCR)-based detection method for Trichoderma harzianum, which causes green mold disease in mushroom cultivation fields and facilities. Based on the sequence data of the internal
transcribed spacer (ITS) region of T. harzianum strains and several other species, six primers consisting of three forward and three reverse primers were designed. Among
the nine possible combinations of these primers, PCR with the pair THITS-F2 and THITS-R3 distinguished most T. harzianum strains from other Trichoderma species. The optimal annealing temperature for detection of T. harzianum strains was from 62° to 63°C with this primer combination. We designed new primers derived from THITS-F2 and THITS-R3. Annealing
temperatures to detect T. harzianum ranged from 64° to 67°C using the new primers. The detection limit of T. harzianum DNA was 50 fg by nested PCR with THITS-F1 and LR1-1 for the first PCR and the new primers for the second PCR. T. harzianum was readily detectable in contaminated cultures of Lentinula edodes by this method. 相似文献
18.
Dwarf bunt of wheat, caused by Tilletia controversa Kühn, is an important international quarantine disease in many countries. The objective of this investigation was to develop a diagnostic molecular marker generated from intersimple sequence repeat (ISSR) for rapid identification of T. controversa. A total of 60 primers were tested by ISSR to detect DNA polymorphisms between T. controversa and related species. The primer ISSR818 generated a polymorphic pattern displaying a 952‐ bp DNA fragment specific for T. controversa. The marker was converted into a sequence characterized amplified region (SCAR), and specific primers (TCKSF2/TCKSR2) were designed for use in a PCR detection assay. Its detection limit was 1 ng of DNA, which could be yielded by 1.1 μg of teliospores in a 25‐ μl PCR. Conclusively, a method to distinguish T. controversa from similar pathogenic fungi has been successfully developed based on the use of a SCAR marker. 相似文献
19.
Specific and Sensitive Detection of Phytophthora nicotianae by Nested PCR and Loop‐mediated Isothermal Amplification Assays
下载免费PDF全文
![点击此处可从《Journal of Phytopathology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Benjin Li Peiqing Liu Shiyong Xie Rongmei Yin Qiyong Weng Qinghe Chen 《Journal of Phytopathology》2015,163(3):185-193
Phytophthora nicotianae is an important soilborne plant pathogen. It causes black shank in tobacco and other commercially important crop diseases. Early and accurate detection of P. nicotianae is essential for controlling these diseases. In this study, primers based on the Ras‐related protein gene (Ypt1) of P. nicotianae were tested for their specific detection of the pathogen using nested PCR and LAMP assays. For specificity testing, DNA extracts from 47 P. nicotianae isolates, 45 isolates of 16 different oomycetes and 25 isolates of other fungal species were used; no cross‐reaction with other pathogens was observed. The sensitivity assay showed that the nested PCR and LAMP assays had detection limits of 100 fg and 10 fg genomic DNA per 25‐μl reaction, respectively. Furthermore, the nested PCR and LAMP assays were used for the detection of DNA from naturally P. nicotianae‐infected tobacco tissues and soil. Our results suggest that the LAMP assay has the greatest potential for the specific detection of P. nicotianae in regions that are at risk of contracting tobacco black shank disease and that the Ypt1 gene is a novel and effective target of P. nicotianae LAMP visual detection. 相似文献
20.
The strain ZK7 of Pochonia chlamydosporia var. chlamydosporia and IPC of Paecilomyces lilacinus are highly effective in the biological control against root-knot nematodes infecting tobacco. When applied, they require
a specific monitoring method to evaluate the colonization and dispersal in soil. In this work, the randomly amplified polymorphic
DNA (RAPD) technique was used to differentiate between the two individual strains and 95 other isolates, including isolates
of the same species and common soil fungi. This approach allowed the selection of specific fragments of 1.2 kb (Vc1200) and
2.0 kb (Vc2000) specific for ZK7, 1.4 kb (P1400) and 0.85 kb (P850) specific for IPC, using the random Primers OPL-02, OPD-05,
OPD-05 and OPC-11, respectively. These fragments were cloned, sequenced, and used to design sequence-characterized amplification
region (SCAR) primers specific for the two strains. In classical polymerase chain reaction (PCR), with serial dilution of
ZK7 and IPC pure culture DNAs template, the detection limits of these oligonucleotide SCAR-PCR primers were found to be 10,
1000, 500, 100 pg, respectively. In the dot blotting, digoxigenin (DIG)-labeled amplicons from these four primers specifically
recognized the corresponding fragments in the DNAs template of these two strains. The detection limit of these amplicons were
0.2, 0.2, 0.5, 0.5 μg, respectively. 相似文献