Diagnosis of PCP by ITSs Nested PCR on Notinvasive Oropharingeal Samples

Pneumocystis carinii pneumonia is an opportunistic disease usually affecting immunocompromised hosts. Diagnosis is based on the microscopical detection of microrganism on BAL, an invasive sample which requires pt compliance for collection and skilled health care personnel for examination. In order to verify the possible diagnostic utility of noninvasive specimens collected in the upper respiratory tract we examined by the Internal Transcribed Spacers (ITSs) nested PCR, 39 oropharingeal samples (garglings) collected from 20 HIVAb positive pts and from 15 healthy controls.     

P. carinii DNA Detected by ITSs Nested PCR in Serum and PBMC of AIDS Patients with PCP

SUMMARY Pneumocystis carinii pneumonia (PCP) is usually diagnosed by examination of BAL, a sample often unpleasant to be collected from immunocompromised host affected by acute respiratory disease. We studied by the Internal Transcribed Spacers (ITSs) nested PCR the presence of P. carinii DNA in serum and Peripheral Blood Mononuclear Cells (PBMC) during acute episodes of PCP to test blood as a possible noninvasive diagnostic tool.     

ITSs Typing of P. carinii Samples from Italy, The Netherlands and Tanzania.

SUMMARY P. carinii molecular epidemiology appears a new interesting investigational field to understand distribution and incidence of isolates from different geographical locations. Recently a typing system, the Type Specific Oligoblotting (TSO) based on 6 different sequences of the Internal Transcribed Spacers (ITSs) of P. carinii rRNA has been developed [1]. By using P. carinii ITSs nested PCR followed by TSO hybridization we have typed 55 lung derived specimens collected in Italy, The Netherlands and sub-Saharian Africa from pts with microwpically detected P. carinii pneumonia.     

Specific PCR-primers as a reliable tool for the detection of white truffles in mycorrhizal roots

During their symbiotic phase, white truffles are barely distinguishable morphologically, and molecular probes are needed for their identification. Here we report the design of species-specific primers for two white truffles ( Tuber magnatum and T. borchii ) on the basis of their ITS sequence. Their efficiency has been successfully tested on fruit bodies of the same species, many related and unrelated fungal species, as well as mycorrhizal roots in direct, nested and multiplex PCR experiments. They have allowed us to identify T. magnatum mycorrhizas for the first time.