Validation of a Real-Time PCR Assay for Identification of Fresh and Processed Carica papaya Botanical Material: Using Synthetic DNA to Supplement Specificity Evaluation.

Several commercially important botanicals have a lack of diagnostic testing options that can quickly and unambiguously identify materials of different matrices. Real-time PCR can be a useful, orthogonal approach to identification for its exceptional specificity and sensitivity. Carica papaya L. is a species with a lack of available identification methods, and one which features two distinct commercially relevant matrices: fresh fruit and powdered fruit extract. In this study, we demonstrate the successful design and validation of a real-time PCR assay for detection of papaya DNA extracted from the two matrices. We also propose a technique that can be used during exclusivity panel construction, when genuine botanical samples are not available for certain species: substitution with synthetic DNA. We demonstrate the use of this material to complete a comprehensive specificity evaluation and confidently determine suitable Ct cutoff values. Further, we demonstrate how ddPCR can be used to determine the copy number of the target sequence in a set amount of genomic DNA, to which synthetic DNA samples can be corrected, and how it can verify specificity of the primers and probe. Through the presentation of successful assay validation for papaya detection, this work serves as a guideline for how to approach specificity evaluation when non-target botanical samples are difficult to obtain and otherwise may not have been included in the exclusivity panel.

Medicaid drug utilization review annual reports for federal fiscal year 1999: looking back to move forward.

OBJECTIVE: To review states' Medicaid drug utilization (DUR) annual reports for federal fiscal year (FFY) 1999 to assess the usefulness of the information provided in assisting states in improving their DUR programs. DATA SOURCES: Medicaid DUR annual reports from 43 states and the District of Columbia. DATA EXTRACTION: Five DUR experts conducted in-depth reviews of the annual reports to determine whether states provided the information required by the Health Care Financing Administration (HCFA; now the Centers for Medicare & Medicaid Services) annual report instructions, to assess the usefulness of information provided by states to other states seeking to improve their programs, and to consider what changes in report content are needed to increase the value of the reports. DATA SYNTHESIS: States provided the information requested by HCFA, but such information was not particularly useful in helping other states improve their programs, nor was it easily available to other states. These reports need to be reoriented. Rather than instruments useful for compliance monitoring, the reports should be remade as tools useful for educating DUR program staff. To do this, changes in report content are needed, and individual states should have access to reports from all states. Specific recommendations made based on these findings would be important first steps in improving the effectiveness of DUR programs. CONCLUSION: Much room for improvement remains in the content and distribution of Medicaid DUR annual reports. Finding ways to improve the effectiveness of DUR is especially important as the nation moves to implement a Medicare prescription drug benefit.