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Objective@#To develop RT-nPCR assays for amplifying partial and complete VP1 genes of human enteroviruses (HEVs) from clinical samples and to contribute to etiological surveillance of HEV-related diseases.@*Methods@#A panel of RT-nPCR assays, consisting of published combined primer pairs for VP1 genes of HEV A-C and in-house designed primers for HEV-D, was established in this study. The sensitivity of each RT-nPCR assay was evaluated with serially diluted virus stocks of five serotypes expressed as CCID @*Results@#The sensitivity of RT-nPCR assays for amplifying partial VP1 gene of HEVs was 0.1 CCID @*Conclusion@#This RT-nPCR system is capable of amplifying the partial and complete VP1 gene of HEV A-D, providing rapid, sensitive, and reliable options for molecular typing and molecular epidemiology of HEVs in clinical specimens.
Asunto(s)
Humanos , Proteínas de la Cápside/genética , Enterovirus Humano A/genética , Enterovirus Humano B/genética , Enterovirus Humano C/genética , Enterovirus Humano D/genética , Epidemiología Molecular/métodos , Tipificación Molecular/métodos , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodosRESUMEN
Purpose: The objective of this study was to develop a sensitive, specific and rapid approach to diagnose hand foot and mouth disease (HFMD) for an early treatment by using loop‑mediated isothermal amplification (LAMP) technique. Materials and Methods: A reverse‑transcription loop‑mediated isothermal amplification (RT‑LAMP) for detecting EV71 virus was developed, the specificity and sensitivity of RT‑LAMP was tested, and the clinical specimens was assayed by the RT‑LAMP comparing with conventional reverse‑transcription polymerase chain reaction (RT‑PCR) and real‑time PCR. Results: A total of 116 clinical specimens from the suspected HFMD individual were detected with the RT‑LAMP. The detection rate for EV71 was 56.89% by RT‑LAMP, 41.38% by real‑time PCR and 34.48% by RT‑PCR. The minimum detection limit of RT‑LAMP was 0.01 PFU, both of RT‑PCR and real‑time PCR was 0.1PFU. Non‑cross‑reactive amplification with other enteroviruses was detected in the survey reports. Conclusions: The effectiveness of RT‑LAMP is higher than RT‑PCR and real‑time PCR. The protocol is easy to operate and time saving. It was not an expensive instrument, which was needed; it is an applicable method for rapid diagnosis of the disease, especially in resource‑poor countries or in developing countries.
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Background: Candida species are now recognized as major causative agents of hospital-acquired infection. One of the major factors contributing to the virulence of Candida is its ability to form surface-attached microbial communities known as "biofilms". The importance of Candida biofilms is because of its increased resistance to antifungal therapy and the ability of cells within biofilms to withstand host immune defenses. Objective: This study was undertaken with the objectives of isolating the Candida species and identifying its virulence factor – the biofilm and to determine the role of biofilm in pathogenicity. Materials and Methods: A cross sectional study was conducted amongst the clinical specimens collected from the critical care wards of a tertiary care Hospital at Navi Mumbai from Jan 2009- Feb 2010. Care was taken to collect the samples before any anti fungal treatment. Candida spp were isolated and identified by standard techniques. Results: Out of total 200 different clinical specimens collected and processed, the most commonly isolated species was C. albicans(61.36 %) along with non albicans like C. parapsilosis (9.1%) C. pseudotropicalis (13.64 %) and C. glabrata (15.9%). Conclusion: The data suggests that the capacity of Candida species to produce biofilm appears to be a reflection of the pathogenic potential of the isolates. Isolates of Candida parapsilosis, Candida pseudotropicalisand Candida glabrata all gave significantly less biofilm growth then C. albicans.
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OBJECTIVE To investigate the nonfermenter distribution and drug-resistance in the clinical specimens,and instruct clinical application of antibiotics reasonably. METHODS Bacteria were cultured,isolated and identified with ATB Expression microbe identification system.Drug-resistance was detected with ATB PSE and statistically analyzed. RESULTS A total of 1 106 strains of bacteria were isolated from 7 395 specimens,the isolation rate was 14.9%.The isolation rate of Pseudomonas aeruginosa was the highest.The isolation rate of nonfermenter was the highest from sputum specimens.Nonfermenter was the most sensitive to imipenem except for Stenotrophomonas maltophilia and Chryseobacterium meningosepticum,after imipenem was cefoperazone/sulbactam. CONCLUSIONS The nonfermenter isolated from different clinical specimens and its drug-resistance are variant.To reduce the production of drug-resistance and control the nosocomical infection,the specimens must be collected and detected timely,the antibiotics are selected according to the results of susceptibility test.