A MULTIPLEX PCR ASSAY FOR THE DIFFERENTIAL DETECTION OF OPISTHORCHIS FELINEUS AND METORCHIS BILIS.

Opisthorchis felineus and Metorchis bilis are two common small worms that parasitize in the gallbladder and bile ducts of the liver of humans and carnivores. These parasites have a severe impact on health and are considered pathogens of serious diseases worldwide, such as cholangiocarcinoma. However, there are still no commercially available molecular diagnostic kits capable of simultaneously detecting these parasites in humans. Therefore, the study aimed to develop a multiplex PCR analysis that will differentially determine these two opisthorchiasis infections in one reaction. Two specific primer pairs for a multiplex polymerase chain reaction (PCR) were designed based on corresponding mitochondrial genome sequences. The multiplex assay detection limit was assessed by serial dilutions of the genomic DNAs of trematode worms examined. Naturally, infected samples of human bile and feces were tested using the developed assay. A multiplex PCR assay was developed based on mitochondrial DNA that accurately and simultaneously identifies two trematode species in one reaction using specific fragment sizes of 307 and 252 bp for O. felineus and M. bilis, respectively. The optimal reaction conditions, specificity, and sensitivity of the multiplex PCR assay were investigated. The lowest DNA concentration detected was 100 pg for M. bilis and O. felineus in a 25µl reaction system. This study provides an efficient tool for the simultaneous detection of O. felineus and M. bilis. The proposed multiplex PCR assay will be potentially useful in epidemiological studies, diagnosis, and treatment of this mixed opisthorchiasis infection.

Randomized clinical trial of two resin-modified glass ionomer materials: 1-year results.

With institutional review board approval, 33 patients who needed restoration of noncarious cervical lesions (NCCL) were enrolled in this study. A total of 92 NCCL were selected and randomly assigned to three groups: (1) Ambar (FGM), a two-step etch-and-rinse adhesive (control), combined with the nanofilled composite resin Filtek Supreme Plus (FSP; 3M ESPE); (2) Fuji II LC (GC America), a traditional resin-modified glass ionomer (RMGIC) restorative material; (3) Ketac Nano (3M ESPE), a nanofilled RMGIC restorative material. Restorations were evaluated at six months and one year using modified United States Public Health Service parameters. At six months after initial placement, 84 restorations (a 91.3% recall rate) were evaluated. At one year, 78 restorations (a 84.8% recall rate) were available for evaluation. The six month and one year overall retention rates were 93.1% and 92.6%, respectively, for Ambar/FSP; 100% and 100%, respectively, for Fuji II LC; and 100% and 100%, respectively, for Ketac Nano with no statistical difference between any pair of groups at each recall. Sensitivity to air decreased for all three adhesive materials from the preoperative to the postoperative stage, but the difference was not statistically significant. For Ambar/FSP, there were no statistical differences for any of the parameters from baseline to six months and from baseline to one year. For Fuji II LC, surface texture worsened significantly from baseline to six months and from baseline to one year. For Ketac Nano, enamel marginal staining increased significantly from baseline to one year and from six months to one year. Marginal adaptation was statistically worse at one year compared with baseline only for Ketac Nano. When parameters were compared for materials at each recall, Ketac Nano resulted in significantly worse color match than any of the other two materials at any evaluation period. At one year, Ketac Nano resulted in significantly worse marginal adaptation than the other two materials and worse marginal staining than Fuji II LC. Surface texture was statistically worse for Fuji II LC compared with the other two materials at all evaluation periods. The one-year retention rate was statistically similar for the three adhesive materials. Nevertheless, enamel marginal deficiencies and color mismatch were more prevalent for Ketac Nano. Surface texture of Fuji II LC restorations deteriorated quickly.

Randomized clinical trial of four adhesion strategies: 18-month results.

STATEMENT OF THE PROBLEM: With Institutional Review Board approval, 39 patients who needed restoration of noncarious cervical lesions (NCCLs) were enrolled in this study. A total of 125 NCCLs were selected and randomly assigned to four groups: 1) a three-step etch-and-rinse adhesive, Adper Scotchbond Multi-Purpose (MP, 3M ESPE, St Paul, MN, USA); 2) a two-step etch-and-rinse adhesive, Adper Single Bond Plus (SB, 3M ESPE); 3) a two-step self-etch adhesive, Adper Scotchbond SE (SE, 3M ESPE); and 4) a one-step self-etch adhesive, Adper Easy Bond (EB, 3M ESPE). A nanofilled composite resin was used for all restorations. Restorations were evaluated at six months and 18 months using modified U.S. Public Health Service (USPHS) parameters. RESULTS: At six months after initial placement, 107 restorations (85.6% recall rate) were evaluated. At 18 months, 94 restorations (75.2% recall rate) were available for evaluation. The 6 mo/18 mo overall retention rates (%) were 100/90.9 for MP; 100/91.7 for SB; 100/90.9 for SE; and 96.4/92.3 for EB with no statistical difference between any pair of groups at each recall. Sensitivity to air decreased significantly for all adhesives from the preoperative to the postoperative stage and was stable thereafter. Interfacial staining did not change statistically from baseline to six months; however, interfacial staining at the enamel margins was statistically worse at 18 months than at baseline for the two self-etch adhesives EB and SE. Marginal adaptation was statistically worse at 18 months compared with baseline only for EB. This tendency was already significant at the six-month recall. CONCLUSION: Although 18-month retention was similar for the different adhesion strategies, enamel marginal deficiencies were more prevalent for the self-etch adhesives.