3) In addition, the detection limit is very low With only two D

3). In addition, the detection limit is very low. With only two DNA copies, it has a higher sensitivity than the currently applied molecular methods, such as semi-nested PCR learn more (10 pg) (Prariyachatigul et al., 2003), PCR enzyme immunoassay (3.2 pg) (Lindsley et al., 2001), PCR hybridization (0.1 pg) (Vanittanakom et

al., 1998) and nested PCR (0.07 pg) (Zeng et al., 2009). The results of P. marneffei detection by LAMP in 23 paraffin wax-embedded clinical samples and 11 bamboo rat tissues were also highly specific. The etiologic agents of the 23 clinical samples were verified previously using culture and sequencing data. Twelve samples were histopathologically positive; all molecular identifications matched with the clinical diagnoses. Samples from penicilliosis and from the natural bamboo rat host were positive with LAMP, whereas all others, including healthy human skin, proved to be negative. Test results were not inhibited by nontarget

DNA. This makes the LAMP technique highly promising for evaluation and application in problematic clinical see more samples such as blood, urine and sputum. In this study, we have proved with the example of P. marneffei that LAMP is a very efficient method for the quick and sensitive identification of fungal pathogens and opportunists. The method can be applied not only to cultures but also to a variety of clinical samples. This can be of great significance to organisms that cause invasive or disseminated infections that are difficult to cultivate from such samples, such as the zygomycete species. A further application may be for detection without isolation of the fungi in the environment. In summary, in the current study, we proved that the LAMP technique enables specific detection of P. marneffei and excludes related biverticillate penicillia and Talaromyces teleomorphs. Similar results were obtained in Paracoccidioides (Endo et al., 2004), Candida (Inacio et al., 2008) and Ochroconis (Ohori et al., 2006). However, in Fonsecaea, identification was possible only at the generic level (Najafzadeh, 2009). An explanation for this phenomenon may be found in the fact that

Penicillium species are relatively distant from each other, with ITS barcoding gaps well over 1%, whereas in Fonsecaea ITS, interspecific Ureohydrolase differences are a few bases only, species delimitations being based on multilocus analyses. We thank Prof. Yokoyama (Center for Pathogenic Fungi and Microbial Toxicoses Chiba University, Chiba, Japan) for providing the reference strains taxonomically close to P. marneffei included in this paper. This study was supported partly by a grant (30770121/2007) from the National Natural Science Foundation of China. “
“Human parturition is associated with an intrauterine pro-inflammatory environment in the choriodecidua. Evidence that some mediators of this signaling cascade also elicit responses leading to labor prompted us to characterize the cellular sources of these mediators in the human choriodecidua.

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