Salivary testing for periodontal disease diagnosis and treatment.

For more than 50 years, clinicians have relied primarily on the same visual and mechanical assessment methods to diagnose and classify periodontal disease. Clinical signs are simply a measurement of the past damage of a disease process. While clinical presentation and probing depths are indicators that the disease exists, these alone cannot determine the types and quantities of the responsible pathogens. Likewise, clinical signs alone cannot determine if therapy has achieved the goal of suppression of the etiological agent(s). Genetic presymptomatic testing complements bacterial DNA testing by providing insight into the patient's genetic predisposition to periodontal disease before symptoms appear, or when disease is already present. DNA-PCR testing of saliva can help the clinician provide an earlier and more specific diagnosis of disease based on causation. Treatment planning is also enhanced, as therapy can be appropriately modified based on both clinical and biological inflammatory factors. Finally, patient communication and case acceptance can be more readily achieved because the test reports elicit a persuasive "seeing is believing" attitude when reviewing test results with patients. Through the use of accurate periodontal charting, medical and dental risk assessments, and other diagnostic screening tests such as OralNA's MyPerioPath and MyPerioID PST tests, highly personalized periodontal therapy can be developed and carried out by the general practitioner. There has never been a better time to become more aware, and keep tighter control, of the periodontal status of one's patient base. Many patients are asking dentists about the connection between periodontal health and general health, While it currently would not be appropriate to suggest a causative relationship, there is abundant ongoing research that suggests a correlative relationship between periodontal disease and other whole-body ailments. Many patients have refused periodontal care or denied the importance of maintaining their periodontal health. The use of tools such as the OralDNA test report can assist in achieving patient acceptance of needed treatment and cooperation with the clinician to improve their periodontal health. The contents of the report and the visual presentation demonstrate that many patients have an active infection that can be stabilized if treated. Patients also provide more information about other factors that may contribute to their periodontal condition. Further, the OralDNA report enhances dentist-physician communication and a team approach to patient care.

Type-specific prevalence and persistence of human papillomavirus in women in the United States who are referred for typing as a component of cervical cancer screening.

OBJECTIVE: The purpose of this study was to report type-specific prevalence and persistence of human papillomavirus (HPV) in women who underwent cytologic screening. STUDY DESIGN: We examined HPV prevalence in 73,371 women who had type-specific HPV testing in 1 of 23 clinical laboratories in the United States. Persistence was evaluated in 963 women who were tested within 8-16 months of their index test. RESULTS: HPV was detected in 31% of the women, and high-risk HPV was detected in 23% of the women. HPV-16, -53, -52, and -31 were the most prevalent types. Of the 953 women with 2 tests, 39% of the women had persistent HPV infection. High-risk HPV persistence was detected in 34% of the women who were positive initially for high-risk HPV. CONCLUSION: Approximately one-third of our sample had HPV; of those women who were retested within 8-16 months, more than one-third had persistent infection. Among women with high-risk HPV infections, the likelihood of persistence was highest with HPV genotypes that were phylogenetically similar to HPV-16.

The diagnosis of Trichomonas vaginalis in liquid-based Pap tests: correlation with PCR.

The conventional Papanicolaou smear (CPS) is not considered accurate for the diagnosis of Trichomonas vaginalis (T. vaginalis), and women noted to carry the organism on CPS are recommended to undergo confirmatory testing. Liquid-based preparations have been shown to facilitate the diagnosis of squamous lesions and may also facilitate the diagnosis of T. vaginalis. We used polymerase chain reaction (PCR) to investigate the accuracy of the diagnosis of T. vaginalis by the liquid-based Pap test (LBP). LBP with the diagnosis of T. vaginalis from a 12-mo period were identified. Residual samples from these cases were subjected to PCR for T. vaginalis as were the residual samples of a control group of 195 LBP (including 103 inflammatory LBP and 69 cases of atypical squamous cells) in which T. vaginalis was not diagnosed cytologically. PCR confirmed the presence of T. vaginalis in 50 of 51 (98%) LBP and identified 2 additional cases. Morphologic identification of T. vaginalis on LBP is highly accurate and should not require confirmatory testing.

The molecular characterization of fatal infectious mononucleosis.

We describe a retrospective study of 4 cases of sporadic fatal infectious mononucleosis (IM), 1 case of fatal IM, and 1 case of sporadic severe IM. Patients were 26 months to 17 years old; 3 were male. Five died of complications of IM. All 5 of these patients had the Epstein-Barr virus (EBV) present in examined tissue specimens; EBV was monoclonal in 3 patients and biclonal in 1. EBV clonality studies were not performed in the remaining patient. All 5 patients also had monoclonal gene rearrangements. The sixth patient survived despite a life-threatening clinical course; EBV was oligoclonal, and gene rearrangements were not detected. EBV clonality and gene rearrangement studies may be usefulfor predicting which patients with clinically aggressive IM are at highest risk for fatal outcome. Patients in whom IM has a fatal outcome are more likely to have monoclonal or biclonal EBV and immunoglobulin heavy chain or T-cell receptor gene rearrangements. In contrast, patients with nonfatal IM may lack monoclonal EBV and monoclonal rearrangements of the aforementioned genes. The reasons EBV induces a monoclonal proliferation only in some patients remain to be elucidated.

Low-density addressable array for the detection and typing of the human papillomavirus.

We have developed a low-density DNA array for the detection and typing of human papillomavirus (HPV) DNA. The gene chemistry strategy involves using a combination of the polymerase chain reaction (PCR) with the consensus oligonucleotide primers MY09/MY11 followed by a ligase detection reaction (LDR). Fluorochrome-labeled HPV-specific primers are joined to a common primer modified with a unique anchoring sequence called a zip code on its 3' end. The result is a series of 60-70 base pair and single-stranded ligation products that are then hybridized to their respective zip code complements affixed to glass slide based arrays. Nine separate zip codes were assigned, one for each HPV type (6,11,16,18, 31, 33, 35, and 53) and one for a beta-globin internal control marker. Two additional zip-codes were reserved for a pair of consensus HPV LDR products: the cLDR1 and cLDR2 primers hybridize to a conserved sequence within the HPV L1 open reading frame internal to the MY09/MY11 fragment. These consensus primers were shown to detect over 40 different HPV types. The purpose of this study was to evaluate the analytic performance of this low-density microarray based assay for HPV, as well as to introduce our simplified read-out instrumentation, shown here to be a low cost and highly efficient way to detect and genotype HPV for clinical testing.

MLL amplification in myeloid malignancies: clinical, molecular, and cytogenetic findings.

Structural rearrangements involving the MLL gene at 11q23 are common recurring abnormalities in de novo and therapy-related hematologic disorders. MLL rearrangement most often results from translocation or partial tandem duplication, although recent published reports suggest a different mechanism by which MLL might participate in leukemogenesis: MLL amplification. We report two patients with myeloid disorders who showed amplification of MLL at diagnosis and who, like the majority of reported cases, had an older age at onset and on aggressive clinical course. Additionally, we summarize the salient clinical, cytogenetic and molecular findings of the 29 other cases of MLL amplification that have thus far been reported.

Clinical and laboratory management of the prothrombin G20210A mutation.

OBJECTIVE: To make recommendations regarding the appropriate evaluation for the prothrombin G20210A mutation, as reflected by published evidence and the consensus opinion of recognized experts in the field. DAT SOURCES: Review of the medical literature, primarily since 1996. DATA EXTRACTION AND SYNTHESIS: After an initial assessment of the literature, key points defining the condition, and review of the clinical study design, a draft manuscript was prepared and circulated to every participant in the College of American Pathologists Conference on Diagnostic Issues in Thrombophilia before the meeting. Each of the key points and associated recommendations were then presented for discussion at the conference. Recommendations were accepted if a consensus of 70% of experts attending the conference was reached. The results of the discussion were used to revise the manuscript into its final form. CONCLUSIONS: Consensus was reached on several recommendations concerning the criteria for testing for the prothrombin G20210A mutation and for the method of testing. First, a major point of consensus was that the prothrombin G20210A mutation is a significant risk factor for venous thromboembolism (VTE) and that testing should be considered in the initial evaluation of suspected inherited thrombophilia. Second, although several analytic methods are commonly used for genetic testing for the prothrombin mutation, all are generally robust and reliable. The recommendations for testing for the prothrombin mutation parallel those for the factor V Leiden mutation and include patients with a history of recurrent VTE, a first episode of VTE before the age of 50 years, a history of an unprovoked VTE at any age, thromboses in unusual anatomic sites, or an affected first-degree relative with VTE. A history of VTE related to pregnancy or estrogen use and unexplained pregnancy loss during the second or third trimesters were also considered to be indications for testing. Other scenarios remain controversial or not recommended, including general population screening.

Hyperhomocyst(e)inemia and Thrombophilia.

OBJECTIVE: To review the role of an elevated total plasma homocysteine level (hyperhomocyst[e]inemia) in patients with venous or arterial thrombosis, as reflected by the medical literature and the consensus opinion of recognized experts in the field. DATA SOURCES: Review of the medical literature, primarily from the last 10 years. DATA EXTRACTION AND SYNTHESIS: The literature was reviewed to identify key points defining the condition, and the clinical study design of each article was examined. A draft manuscript was prepared and circulated prior to the conference to every participant in the College of American Pathologists Conference XXXVI: Diagnostic Issues in Thrombophilia. Each of the key points and associated recommendations was then presented for discussion at the conference. Recommendations were accepted if a consensus of the 70% of the experts attending the conference was reached. The results of the discussion were used to revise the manuscript into its final form. CONCLUSIONS: Consensus was reached on 9 recommendations concerning the criteria for diagnosis, the method of testing, and the approach for clinical management. A major point of consensus was that no causal role of hyperhomocyst(e)inemia in venous or arterial thrombosis is yet established. Testing methods used to measure homocysteine directly are sensitive and reliable, and provide more information than does genotyping for markers linked to abnormal plasma homocysteine.

Protein self-organization patterns in dried serum reveal changes in B-cell disorders.

BACKGROUND: Detection of serum monoclonal proteins is a common laboratory analysis used in the evaluation of patients with B-cell disorders. Since many individuals with elevated immunoglobulin have no symptoms, it is important to have simple methods for initial screening of patients with suspected B-cell disorders. METHODS: Samples of serum from healthy donors and from patients with elevated immunoglobulin levels were tested using a technology named Droplet MicroChromatography (DMC). DMC was developed at Artann Laboratories (West Trenton, New Jersey, USA) for the rapid assessment of changes in the composition of serum. DMC is based on the dynamics of the sediment pattern formation during drying of a fluid microdroplet. RESULTS: Results of this pilot study confirm the hypothesis that the pattern formation created by drying droplets of serum would differ between normal samples and those containing monoclonal proteins. Reproducible differences in the patterns formed by the two types of specimens are shown. Strong correlation between abnormally elevated levels of immunoglobulins in the serum of myeloma patients and the patterns formed by drying droplets of serum indicates that the DMC technique may be suitable for semi-quantitative analysis of serum samples. We also demonstrate that computer identification of the drying droplet structure and dynamics is a tractable issue. CONCLUSIONS: DMC has significant diagnostic potential and can serve as a basis for development of a simple, rapid, and inexpensive method for initial screening of patients suspected of having multiple myeloma and other pathologies of lymphoid origin that are associated with the overproduction of monoclonal immunoglobulins. The DMC test requires only approximately 1 microL of serum and could therefore be performed in any facility where it is safe to work with serum.