Janet G. Baseman

Diagnostic Assessment of Mycoplasma genitalium in Culture-Positive Women

ABSTRACT Detection of Mycoplasma genitalium-mediated, chlamydia-negative nongonococcal urethritis and other M. genitalium-linked infectious etiologies has been very challenging. Although M. genitalium is considered a leading cause of genitourinary symptoms in men and women, extreme difficulties in its cultivation due to its highly fastidious nature and the lack of routine and effective diagnostic tests have slowed the generation of clinical data which directly implicate the presence of M. genitalium in disease pathogenesis. In this study, we compared enzyme-linked immunosorbent assays (ELISAs) and immunoblot and PCR assays in M. genitalium culture-positive women over 1 to 3 years of clinical visits to determine the usefulness of independent diagnostic strategies. Furthermore, the value of combinatorial diagnostic assessments is described, which provides insights into the dynamics of M. genitalium-host interactions. Overall, we show that neither ELISA nor PCR, alone or in combination, provides the sensitivity required to confidently predict the existence of viable M. genitalium organisms in cervical and vaginal samples. Additionally, culture-positive women exhibited a range of antibody responsiveness to M. genitalium based upon ELISA and immunoblot assessments, indicating immune diversity among this high-risk population.

Determination of Infectious Load of Mycoplasma genitalium in Clinical Samples of Human Vaginal Cells

ABSTRACT Mycoplasma genitalium is a leading cause of chlamydia-negative, nongonoccocal urethritis and has been directly implicated in numerous other genitourinary as well as extragenitourinary tract pathologies. Detection of M. genitalium has relied almost entirely on PCR amplification of clinical specimens and evidence of seroconversion since these mycoplasmas are highly fastidious and culture isolation by microbiological techniques is very rare. We have established a combinatorial strategy using confocal immunoanalysis (CIA) and real-time PCR to qualitatively and quantitatively assess patterns of M. genitalium infection in women attending a sexually transmitted disease-related health clinic in San Antonio, Tex. CIA allows spatial examination of mycoplasmas on surfaces and inside human target cells, plus the ability to evaluate cell-to-cell patterns and variances within samples. Real-time PCR permits determination of genome copy numbers of mycoplasmas and human cells by multiplex amplification using mycoplasma gyrA and human RNase P gene sequences, which indicates overall levels of mycoplasma infection and degree of parasitism. These assays are strongly correlated and, in combination, permit detection and elucidation of heretofore-unrecognized patterns of M. genitalium infections in clinical and experimental samples.

Mycoplasma genitalium symptoms, concordance and treatment in high-risk sexual dyads.

The objective of this study was to determine the prevalence and concordance of Mycoplasma genitalium (MG) among Mexican American and African American women and their male sexual partners. Secondary objectives were to determine symptoms of MG infection and persistence of MG after antibiotic therapy. Heterosexual couples were tested for MG and interviewed separately regarding symptoms and behavioural/epidemiologic variables at baseline, six and 12 months. The overall prevalence of MG among women and men was 9.5% and 10.6%, respectively. Subjects were five times more likely to be infected with MG if their sexual partner was MG positive. Among men and women, MG prevalence and mean bacterial loads were similar after receiving single-dose azithromycin, doxycycline or no antibiotics. MG was associated with current urethral discharge in men. No clinical symptoms were specifically diagnostic of MG infection in women.

Persistence of Community-Acquired Respiratory Distress Syndrome Toxin-Producing Mycoplasma pneumoniae in Refractory Asthma

BACKGROUND The role of Mycoplasma pneumoniae (Mp) in the initiation and persistence of asthma remains elusive. Mp community-acquired respiratory distress syndrome toxin (CARDS Tx) is a unique virulence factor that induces an intense lymphocytic response and exacerbates asthma in animal models. We sought to determine the incidence of Mp infection and the presence of CARDS Tx in subjects with refractory asthma (RA). METHODS We conducted a prospective observational study in 64 subjects with RA. Respiratory secretions (sputum, nasal lavage, and throat swab) and blood were analyzed for the presence of CARDS Tx and P1 adhesin (P1) DNA by polymerase chain reaction (PCR), and CARDS Tx by antigen capture. Serum IgM and IgG antibodies to CARDS Tx were determined by enzyme-linked immunosorbent assay (ELISA). RESULTS Thirty-three of 64 subjects (52%) tested positive for Mp: 29 of 33 by CARDS Tx vs 10 of 33 by P1 assays. Ten subjects followed longitudinally for up to 633 days tested persistently positive for Mp. There were no significant differences in Mp-specific IgG responses between Mp-positive and Mp-negative groups. Eight of 10 subjects who tested persistently positive failed to mount a substantial IgG response to CARDS Tx, and up to 8 weeks of clarithromycin failed to eradicate Mp in five subjects. CONCLUSIONS Subjects with RA may be chronically infected with Mp. PCR for CARDS Tx appears to be the most sensitive method of identifying Mp infection. Despite the persistence of Mp in subjects with RA, some subjects failed to mount an IgG response, and macrolide therapy was insufficient to eradicate Mp.

Public health communications and alert fatigue

BackgroundHealth care providers play a significant role in large scale health emergency planning, detection, response, recovery and communication with the public. The effectiveness of health care providers in emergency preparedness and response roles depends, in part, on public health agencies communicating information in a way that maximizes the likelihood that the message is delivered, received, deemed credible and, when appropriate, acted on. However, during an emergency, health care providers can become inundated with alerts and advisories through numerous national, state, local and professional communication channels. We conducted an alert fatigue study as a sub-study of a larger randomized controlled trial which aimed to identify the most effective methods of communicating public health messages between public health agencies and providers. We report an analysis of the effects of public health message volume/frequency on recall of specific message content and effect of rate of message communications on health care provider alert fatigue.MethodsHealth care providers enrolled in the larger study (n=528) were randomized to receive public health messages via email, fax, short message service (SMS or cell phone text messaging) or to a control group that did not receive messages. For 12 months, study messages based on real events of public health significance were sent quarterly with follow-up telephone interviews regarding message receipt and topic recall conducted 5–10 days after the message delivery date. During a pandemic when numerous messages are sent, alert fatigue may impact ability to recall whether a specific message has been received due to the “noise” created by the higher number of messages. To determine the impact of “noise” when study messages were sent, we compared health care provider recall of the study message topic to the number of local public health messages sent to health care providers.ResultsWe calculated the mean number of messages that each provider received from local public health during the time period around each study message and provider recall of study message content. We found that recall rates were inversely proportional to the mean number of messages received per week: Every increase of one local public health message per week resulted in a statistically significant 41.2% decrease (p < 0.01), 95% CI [0.39, .87] in the odds of recalling the content of the study message.ConclusionsTo our knowledge, this is the first study to document the effects of alert fatigue on health care providers’ recall of information. Our results suggest that information delivered too frequently and/or repetitively through numerous communication channels may have a negative effect on the ability of health care providers to effectively recall emergency information. Keeping health care providers and other first-line responders informed during an emergency is critical. Better coordination between organizations disseminating alerts, advisories and other messages may improve the ability of health care providers to recall public health emergency messages, potentially impacting effective response to public health emergency messages.

Implementation and expansion of an electronic medical record for HIV care and treatment in Haiti: An assessment of system use and the impact of large-scale disruptions

PURPOSE System use is a key criterion of success in an electronic medical record (EMR) implementation, and there is little research on long-term use of systems following implementation. The aim of the paper was to describe the development, implementation and use of iSanté, Haitis national HIV care and treatment EMR. METHODS To build a picture of the history of iSanté, we interviewed 11 staff involved with the development and implementation of the EMR, and reviewed organization records. Data entry and report use were ascertained by querying the central patient database. RESULTS By the end of 2010 there were 67 sites with iSanté installed, and the scope of the system had been expanded to include primary care and obstetrics and gynecology. New functionality includes data forms specific to subpopulations, the ability to transfer patient records among clinics, and integration with an electronic laboratory system. We observed fluctuations in use over time, with substantial reductions in the number of active sites during times of large-scale disruptions in Haiti. A surge in report use following the January 2010 earthquake suggests that clinics found the EMR to be a valuable source of data during the recovery phase. CONCLUSION There is real potential for EMRs in developing countries to improve clinical practice and make data available for efficient reporting, quality improvement and other population health uses. An approach of continuous system improvement, combined with regular assessments of use, is necessary for achieving an effective, national implementation of a standardized EMR. We have achieved successes in terms of rolling out new functionality and expanding to new sites, but more work remains to be done to improve perceptions of data quality and increase use of population data for accurate and timely reporting.

Effect of 6F urethral catheterization on urinary flow rates during repeated pressure-flow studies in healthy female volunteers.

OBJECTIVES To prospectively examine the effect of a 6F urethral catheter on the urinary flow rate in healthy women without lower urinary tract symptoms undergoing pressure-flow studies. METHODS Women between the ages of 30 and 70 years, without lower urinary tract complaints and without a history of surgery for incontinence, were recruited. All women completed the Urogenital Distress Inventory-6 questionnaire, and only women with mild or no symptoms were enrolled. After a free flow rate was determined, cystometry and pressure-flow studies were performed once and then repeated using a 6F urethral catheter. The peak flow rates during the first and second studies were compared with one another and with the nonintubated values. RESULTS Of the 59 women that applied, 20 volunteers (mean age 41.7 years) met the entry criteria and were evaluated. The mean nonintubated flow rate was 22.65 mL/s. With a 6F urethral catheter in place, our cohort had a mean peak flow rate of 16.25 mL/s on the first study and 15.25 mL/s on the second. A significant difference was demonstrated between the free and intubated peak flow rates for both the first (P = 0.0006) and the second (P = 0.0001) study. No significant difference was detected between the two intubated peak flow rates (P = 0.262). CONCLUSIONS Our data suggest that the presence of a 6F urethral catheter significantly reduces the maximal flow rate compared with free flow studies. Whether these findings represent a truly obstructive effect of the catheter, detrusor fatigue, or other possible effect of the catheter remains to be elucidated.

Evaluation of primary cervical cancer screening with an oncogenic human papillomavirus DNA test and cervical cytologic findings among women who attended family planning clinics in the United States.

OBJECTIVE Our goal was to evaluate the performance of screening with (1) Papanicolaou and human papillomavirus (HPV) DNA testing and (2) Papanicolaou testing with reflex HPV testing of atypical squamous cells of undetermined significance for detecting cervical intraepithelial neoplasia grade 3 or more in clinics that serve low-income women in the United States. STUDY DESIGN There were 4799 women who were recruited primarily from Planned Parenthood clinics and who were screened with liquid-based Papanicolaou testing and HPV DNA testing and referred for biopsy based on a positive test result for oncogenic HPV DNA or a Papanicolaou test that showed atypical squamous cells of undetermined significance or more. RESULTS Among 931 women who were 30-50 years of age, the sensitivity of reflex HPV testing was 53.8% (range, 38.2%-72.3%). The sensitivity of HPV DNA and Papanicolaou testing was 91% (range, 74.6%-100%). The specificity of reflex HPV testing was 95.1% (range, 93.8%-96.3%). Generally, the specificity of HPV DNA and Papanicolaou testing was low. CONCLUSION Among US women who are >or=30 years old, HPV DNA and Papanicolaou testing is a reasonable cervical cancer screening strategy.

Public Health Practice within a Health Information Exchange: Information Needs and Barriers to Disease Surveillance

Introduction: Public health professionals engage in frequent exchange of health information while pursuing the objectives of protecting and improving population health. Yet, there has been little study of the information work of public health workers with regard to information exchange. Our objective was to gain a better understanding of information work at a local health jurisdiction before and during the early stages of participation in a regional Health Information Exchange. Methods: We investigated the information work of public health workers engaged in disease surveillance activities at a medium-sized local health jurisdiction by conducting semi-structured interviews and thematically analyzing interview transcripts. Results: Analysis of the information work of public health workers revealed barriers in the following areas: information system usability; data timeliness, accuracy and completeness; and social interaction with clients. We illustrate these barriers by focusing on the work of epidemiologists. Conclusion: Characterizing information work and barriers to information exchange for public health workers should be part of early system design efforts. A comprehensive understanding of the information practice of public health workers will inform the design of systems that better support public health work.

Sequence Variation of Human Papillomavirus Type 16 and Measurement of Viral Integration by Quantitative PCR

Given that the integration of human papillomavirus type 16 (HPV16) into the host genome occurs preferentially with the disruption of the E2 gene, a ratio of E2 to E7 gene copies is often used as a marker for integration. It is largely undetermined, however, whether ratio estimates are affected by HPV intratypic variations. We assembled four plasmid constructs, each containing a DNA fragment from an HPV16 European, Asian-American, African-1, or African-2 variant. These constructs and nine cervical swab samples were assayed by real-time PCR with two primer-probe sets for each gene: a specific set, fully complementary to the HPV16 prototype, and a degenerate set, incorporating degenerate bases at positions where nucleotides differed among the variants. The ratio of E2 to E7 gene copies for the European variant construct was close to 1, no matter which sets of primers and probes were used. While the ratios for the African-1 and Asian-American variant constructs remained close to 1 with the degenerate sets of primers and probes, the ratios were 0.36 and 2.57, respectively, with the specific sets of primers and probes. In addition, a nucleotide alteration at the position immediately following the 3′ end of the E2 forward primer binding site was found to be responsible for an underestimation of E2 gene copies for the African-2 variant construct. Similar patterns were found in nine cervical samples. In conclusion, mismatches between the primers and probes and their targets due to HPV16 intratypic variations would introduce errors in testing for integration; this situation can be sufficiently ameliorated by incorporating degenerate bases into the primers and probes.