Rapid diagnostic tests as a source of DNA for Plasmodium species-specific real-time PCR

<p>Abstract</p> <p>Background</p> <p>This study describes the use of malaria rapid diagnostic tests (RDTs) as a source of DNA for <it>Plasmodium </it>species-specific real-time PCR.</p> <p>Methods</p> <p>First, the best method to recover DNA from RDTs was investigated and then the applicability of this DNA extraction method was assessed on 12 different RDT brands. Finally, two RDT brands (OptiMAL Rapid Malaria Test and SDFK60 malaria Ag <it>Plasmodium falciparum</it>/Pan test) were comprehensively evaluated on a panel of clinical samples submitted for routine malaria diagnosis at ITM. DNA amplification was done with the 18S rRNA real-time PCR targeting the four <it>Plasmodium </it>species. Results of PCR on RDT were compared to those obtained by PCR on whole blood samples.</p> <p>Results</p> <p>Best results were obtained by isolating DNA from the proximal part of the nitrocellulose component of the RDT strip with a simple DNA elution method. The PCR on RDT showed a detection limit of 0.02 asexual parasites/μl, which was identical to the same PCR on whole blood. For all 12 RDT brands tested, DNA was detected except for one brand when a low parasite density sample was applied. In RDTs with a plastic seal covering the nitrocellulose strip, DNA extraction was hampered. PCR analysis on clinical RDT samples demonstrated correct identification for single species infections for all RDT samples with asexual parasites of <it>P. falciparum </it>(n = 60), <it>Plasmodium vivax </it>(n = 10), <it>Plasmodium ovale </it>(n = 10) and <it>Plasmodium malariae </it>(n = 10). Samples with only gametocytes were detected in all OptiMAL and in 10 of the 11 SDFK60 tests. None of the negative samples (n = 20) gave a signal by PCR on RDT. With PCR on RDT, higher Ct-values were observed than with PCR on whole blood, with a mean difference of 2.68 for OptiMAL and 3.53 for SDFK60. Mixed infections were correctly identified with PCR on RDT in 4/5 OptiMAL tests and 2/5 SDFK60 tests.</p> <p>Conclusions</p> <p>RDTs are a reliable source of DNA for <it>Plasmodium </it>real-time PCR. This study demonstrates the best method of RDT fragment sampling for a wide range of RDT brands in combination with a simple and low cost extraction method, allowing RDT quality control.</p

In vitro studies on the release of isoniazid incorporated in poly(epsilon-caprolactone)

A polymeric micro- and nanosphere formulation using poly (E-caprolactone) (PCL) to entrap an antituberculosis drug, isoniazid (INH), was developed and characterized. The microspheres were prepared by a solvent evaporation method using ethyl acetate, PCL and INH as the organic phase and water and Tween 40 as the aqueous phase. The nanospheres were prepared by a spontaneous emulsification solvent diffusion method using 40% ethanol in acetone (v/v), PCL and INH as the organic phase and water and Tween 40 as the aqueous phase. After freeze-drying, these systems were characterized by scanning electron microscopy (SEM), particle size analysis, determination of entrapped INH content, in vitro INH release and brine shrimp toxicity bioassay.18547347

Allergic reaction after rubber dam placement

In the last few years allergic reactions to natural rubber latex (NRL) have increased in dental practice affecting both the dental team and patients. Some case reports discuss the potential risks of hypersensitivity to NRL products. An adverse patient reaction after dental rubber dam placement is reported. About 1 min after the isolation of the tooth with a rubber dam the patient presented signs and symptoms of hypersensitivity. Oxygen and intravenous hydrocortisone were administered and the patient kept under observation, After 2 h she had stable vital signs and no more allergies symptoms. It is unclear whether components of the NRL dam or the cornstarch powder incorporated with the rubber dam was responsible for the allergic reaction. Dentists must be aware of the health problem and be prepared for an adequate management in dental practice.26318218