Ability To Serologically Confirm Recent Zika Virus Infection in Areas with Varying Past Incidence of Dengue Virus Infection in the United States and U.S. Territories in 2016.

Cross-reactivity within flavivirus antibody assays, produced by shared epitopes in the envelope proteins, can complicate the serological diagnosis of Zika virus (ZIKAV) infection. We assessed the utility of the plaque reduction neutralization test (PRNT) to confirm recent ZIKAV infections and rule out misleading positive immunoglobulin M (IgM) results in areas with various levels of past dengue virus (DENV) infection incidence. We reviewed PRNT results of sera collected for diagnosis of ZIKAV infection from 1 January through 31 August 2016 with positive ZIKAV IgM results, and ZIKAV and DENV PRNTs were performed. PRNT result interpretations included ZIKAV, unspecified flavivirus, DENV infection, or negative. For this analysis, ZIKAV IgM was considered false positive for samples interpreted as a DENV infection or negative. In U.S. states, 208 (27%) of 759 IgM-positive results were confirmed to be ZIKAV compared to 11 (21%) of 52 in the U.S. Virgin Islands (USVI), 15 (15%) of 103 in American Samoa, and 13 (11%) of 123 in Puerto Rico. In American Samoa and Puerto Rico, more than 80% of IgM-positive results were unspecified flavivirus infections. The false-positivity rate was 27% in U.S. states, 18% in the USVI, 2% in American Samoa, and 6% in Puerto Rico. In U.S. states, the PRNT provided a virus-specific diagnosis or ruled out infection in the majority of IgM-positive samples. Almost a third of ZIKAV IgM-positive results were not confirmed; therefore, providers and patients must understand that IgM results are preliminary. In territories with historically higher rates of DENV transmission, the PRNT usually could not differentiate between ZIKAV and DENV infections.

Cluster of Neisseria gonorrhoeae Isolates With High-level Azithromycin Resistance and Decreased Ceftriaxone Susceptibility, Hawaii, 2016.

BACKGROUND: The Centers for Disease Control and Prevention (CDC) currently recommends dual therapy with ceftriaxone and azithromycin for gonorrhea to ensure effective treatment and slow emergence of antimicrobial resistance. Since 2013, the prevalence of reduced azithromycin susceptibility increased in the United States; however, these strains were highly susceptible to cephalosporins. We identified a cluster of Neisseria gonorrhoeae isolates with high-level azithromycin resistance, several of which also demonstrated decreased ceftriaxone susceptibility. METHODS: Eight N. gonorrhoeae isolates collected from 7 patients on Oahu, Hawaii, seen 21 April 2016 through 10 May 2016 underwent routine Etest antimicrobial susceptibility testing by the Hawaii Department of Health. All demonstrated elevated azithromycin minimum inhibitory concentrations (MICs) >256 µg/mL and elevated ceftriaxone MICs (≥0.125 µg/mL). Isolates were sent to the University of Washington and CDC for confirmatory agar dilution testing; sequence data were sent to CDC for analysis. All patients were interviewed and treated, and when possible, partners were interviewed, tested, and treated. RESULTS: All isolates had azithromycin MICs >16 µg/mL and 5 had ceftriaxone MICs = 0.125 µg/mL by agar dilution. All isolates were ß-lactamase positive and were resistant to penicillin, tetracycline, and ciprofloxacin. Genomic analysis revealed genetic relatedness. No patients reported recent travel or antibiotic use, and no male patients reported male sex partners. All patients were successfully treated. CONCLUSIONS: This cluster of genetically related gonococcal isolates with decreased ceftriaxone susceptibility and high-level azithromycin resistance may bring the threat of treatment failure in the United States with the current recommended dual therapy one step closer.

Azithromycin Resistance and Decreased Ceftriaxone Susceptibility in Neisseria gonorrhoeae, Hawaii, USA.

During 2016, eight Neisseria gonorrhoeae isolates from 7 patients in Hawaii were resistant to azithromycin; 5 had decreased in vitro susceptibility to ceftriaxone. Genomic analysis demonstrated a distinct phylogenetic clade when compared with local contemporary strains. Continued evolution and widespread transmission of these strains might challenge the effectiveness of current therapeutic options.

Notes from the Field: Outbreak of Locally Acquired Cases of Dengue Fever--Hawaii, 2015.

On October 21, 2015, the Hawaii Department of Health (HDOH) was notified of a positive dengue immunoglobulin M (IgM) antibody result in a woman residing on Hawaii Island (also known as the Big Island). The patient had no history of travel off the island, and other family members reported having similar signs and symptoms, which consisted of fever, headache, myalgias and arthralgias, and a generalized erythematous rash. HDOH initiated an investigation to identify any additional cases and potential exposure sources. On October 24, HDOH received report of a group of mainland U.S. visitors who had traveled together on Hawaii Island, including several who had developed a febrile illness. Additionally, on October 27, HDOH was notified of an unrelated person, also on Hawaii Island, with a positive dengue IgM result. As of November 26, 2015, HDOH had identified 107 laboratory-confirmed cases of dengue fever, with dates of onset ranging from September 11 to November 18, 2015.

Notes from the Field: Outbreak of Zika Virus Disease - American Samoa, 2016.

During December 2015-January 2016, the American Samoa Department of Health (ASDoH) detected through surveillance an increase in the number of cases of acute febrile rash illness. Concurrently, a case of laboratory-confirmed Zika virus infection, a mosquito-borne flavivirus infection documented to cause microcephaly and other severe brain defects in some infants born to women infected during pregnancy (1,2) was reported in a traveler returning to New Zealand from American Samoa. In the absence of local laboratory capacity to test for Zika virus, ASDoH initiated arboviral disease control measures, including public education and vector source reduction campaigns. On February 1, CDC staff members were deployed to American Samoa to assist ASDoH with testing and surveillance efforts.

Culture-negative prosthetic valve endocarditis with concomitant septicemia due to a nontoxigenic Corynebacterium diphtheriae biotype gravis isolate in a patient with multiple risk factors.

A 54-year-old female with a prosthetic mitral valve presented with a 3-day history of dizziness, subjective fever, and chills. Blood cultures were positive for a pleomorphic Gram-positive rod. Initial phenotypic testing could only support the identification of a Corynebacterium species. Nucleic acid sequencing (16S rRNA) and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) were conclusive for Corynebacterium diphtheriae. Definitive phenotypic testing classified the strain as nontoxigenic C. diphtheriae biotype Gravis.

Gonococcal susceptibility to cephalosporins--Hawaii, 2003 to 2011.

Among gonococcal isolates examined at the Hawaii State Laboratory Division from 2003 to 2011, the prevalence of elevated cefixime minimum inhibitory concentrations (MICs; ≥0.064 µg/mL) and elevated cefpodoxime MICs (≥0.19 µg/mL) increased over time. In contrast, few isolates exhibited elevated ceftriaxone MICs (≥0.094 µg/mL), and the prevalence of elevated ceftriaxone MICs did not change.

Neisseria gonorrhoeae with high-level resistance to azithromycin: case report of the first isolate identified in the United States.

We report on the first Neisseria gonorrhoeae isolate in the United States identified with high-level resistance to azithromycin. This report discusses the epidemiologic case investigation, the molecular studies of resistance-associated mutations and N. gonorrhoeae multiantigen sequence typing, and challenges posed by emerging gonococcal antimicrobial resistance.

Workshop on research priorities for management and treatment of angiostrongyliasis(1).

In a concluding session of the workshop, the participants developed a list of 115 research and outreach needs, outlining the top 5-7 needs in each of 8 areas (Table). For complete information, including presenter details and abstracts, visit the workshop website at www.hawaii.edu/cowielab/Angio%20website%20home.htm.

Pandemic preparedness in Hawaii: a multicenter verification of real-time RT-PCR for the direct detection of influenza virus types A and B.

OBJECTIVE: We integrated multicenter, real-time (RTi) reverse transcription polymerase chain reaction (RT-PCR) screening into a statewide laboratory algorithm for influenza surveillance and response. METHODS: Each of three sites developed its own testing strategy and was challenged with one randomized and blinded panel of 50 specimens previously tested for respiratory viruses. Following testing, each participating laboratory reported its results to the Hawaii State Department of Health, State Laboratories Division for evaluation and possible discrepant analysis. RESULTS: Two of three laboratories reported a 100% sensitivity and specificity, resulting in a 100% positive predictive value and a 100% negative predictive value (NPV) for influenza type A. The third laboratory showed a 71% sensitivity for influenza type A (83% NPV) with 100% specificity. All three laboratories were 100% sensitive and specific for the detection of influenza type B. Discrepant analysis indicated that the lack of sensitivity experienced by the third laboratory may have been due to the analyte-specific reagent probe used by that laboratory. Use of a newer version of the product with a secondary panel of 20 specimens resulted in a sensitivity and specificity of 100%. CONCLUSIONS: All three laboratories successfully verified their ability to conduct clinical testing for influenza using diverse nucleic acid extraction and RTi RT-PCR platforms. Successful completion of the verification by all collaborating laboratories paved the way for the integration of those facilities into a statewide laboratory algorithm for influenza surveillance and response.

Pulmonary Coccidioidomycosis Mimicking Aspergillosis Fungus Ball.

The genus Coccidioides is composed of C. immitis and C. posadasii. Both can cause coccidioidomycosis and are geographically restricted to certain areas of endemicity. The histopathologic features in pulmonary coccidioidomycosis include necrotizing granulomatous inflammation and the presence of spherules, which is considered to be a key diagnostic finding. Cavitary lung disease containing a fungal ball with branching septate hyphae is an unusual funding in pulmonary coccidioidomycosis but is typical for aspergillosis. We present a case of 42 year old man who underwent wedge resection of the lung for a persistent cavitary lesion. The microscopic examination shows a fungal ball composed of acute-angle branching septate hyphae, consistent with a diagnosis of aspergillosis. However, cultures and molecular testing by DNA sequencing of the 28S ribosomal DNA gene confirmed the identification of C. posadasii. This finding highlights the importance of exposure history and organism identification by either conventional cultivation or molecular testing in rendering an accurate diagnosis.

Investigation and Response to an Outbreak of Dengue: Island of Hawaii, 2015-2016.

OBJECTIVES: From September 2015 through March 2016, Hawaii had the largest outbreak of locally transmitted dengue since 1944. We report on the Hawaii Department of Health's (HDOH's) investigation, findings, and response to the outbreak. METHODS: We defined cases of dengue using a modified version of the Council of State and Territorial Epidemiologists' case definition for dengue virus infections. We conducted epidemiologic investigations, including interviews with case-persons, review of medical records, laboratory testing, genetic sequencing of specimens, and geographic information system (GIS) data analysis. Outbreak response included community outreach and vector-control activities. RESULTS: We identified 264 confirmed cases of dengue; illness onset dates ranged from September 11, 2015, to March 17, 2016, all with reported travel to or residence on the Island of Hawaii. Of 264 persons with confirmed dengue, 238 (90.2%) were Hawaii residents. Thirty-seven (14.0%) persons required hospitalization; no cases of severe dengue or death were reported. GIS hot-spot analysis identified a cluster of cases on the western side of the island. Established risk factors for dengue exposure included holes in window or door screens, presence of standing water, and not using insect repellent or wearing protective clothing. CONCLUSIONS: To prevent or mitigate the spread of future arboviral introductions and outbreaks, the public health response should focus on behavioral and cultural attitudes, emphasizing personal mosquito protection and mosquito control at the community level. Outbreak responses can also be enhanced through the use of advanced GIS techniques, such as hot-spot analysis, to provide situational awareness and guide response efforts.

Real-Time Polymerase Chain Reaction Detection of Angiostrongylus cantonensis DNA in Cerebrospinal Fluid from Patients with Eosinophilic Meningitis.

Angiostrongylus cantonensis is the most common infectious cause of eosinophilic meningitis. Timely diagnosis of these infections is difficult, partly because reliable laboratory diagnostic methods are unavailable. The aim of this study was to evaluate the usefulness of a real-time polymerase chain reaction (PCR) assay for the detection of A. cantonensis DNA in human cerebrospinal fluid (CSF) specimens. A total of 49 CSF specimens from 33 patients with eosinophilic meningitis were included: A. cantonensis DNA was detected in 32 CSF specimens, from 22 patients. Four patients had intermittently positive and negative real-time PCR results on subsequent samples, indicating that the level of A. cantonensis DNA present in CSF may fluctuate during the course of the illness. Immunodiagnosis and/or supplemental PCR testing supported the real-time PCR findings for 30 patients. On the basis of these observations, this real-time PCR assay can be useful to detect A. cantonensis in the CSF from patients with eosinophilic meningitis.

Public health laboratory workforce outreach in Hawai'i: CLIA-focused student internship pilot program at the state laboratories.

Chronically understaffed public health laboratories depend on a decreasing number of employees who must assume broader responsibilities in order to sustain essential functions for the many clients the laboratories support. Prospective scientists considering a career in public health are often not aware of the requirements associated with working in a laboratory regulated by the Clinical Laboratory Improvement Amendments (CLIA). The purpose of this pilot internship was two-fold; introduce students to operations in a regulated laboratory early enough in their academics so that they could make good career decisions, and evaluate internship methodology as one possible solution to workforce shortages. Four interns were recruited from three different local universities, and were paired with an experienced State Laboratories Division (SLD) staff mentor. Students performed tasks that demonstrated the importance of CLIA regulations for 10-15 hours per week over a 14 week period. Students also attended several directed group sessions on regulatory lab practice and quality systems. Both interns and mentors were surveyed periodically during the semester. Surveys of mentors and interns indicated overall positive experiences. One-on-one pairing of experienced public health professionals and students seems to be a mutually beneficial arrangement. Interns reported that they would participate if the internship was lower paid, unpaid, or for credit only. The internship appeared to be an effective tool to expose students to employment in CLIA-regulated laboratories, and potentially help address public health laboratory staffing shortfalls. Longer term follow up with multiple classes of interns may provide a more informed assessment.

The current status of laboratory diagnosis of Angiostrongylus cantonensis infections in humans using serologic and molecular methods.

Laboratory diagnosis of angiostrongyliasis relies on serological techniques, since definitive diagnosis is insensitive. Modern antibody detection methods focus on antibodies to the 29 and 31 kDa proteins of the parasite. Antigen detection may ultimately prove to be more reliable than antibody detection but no method has been adopted for clinical diagnostic use. Diagnosis using PCR amplification of DNA sequences specific to Angiostrongylus cantonensis have been developed but have not yet been validated for clinical use. Diagnostic tests have not been developed commercially and in the United States tests developed experimentally by non-commercial laboratories have to be approved by the Food and Drug Administration before they can be sold to other laboratories for diagnostic purposes.

Pacific region influenza surveillance for oseltamivir resistance.

BACKGROUND: Hawaii and the United States-affiliated Pacific islands (USAPI) host over 8 million travelers annually, most of whom originate in Asia, Australia, and the Americas where prevalence of oseltamivir resistance in 2009 pandemic influenza A (H1N1) has been reported to be 2.5-3.5%. OBJECTIVE: To survey a collection of samples from Hawaii and the USAPI that had tested positive for the 2009 pandemic influenza A (H1N1) virus by RTI-PCR to assess whether antiviral resistance emerged in these island communities during the 2009 H1N1 pandemic. STUDY DESIGN: We examined RNA extracted from Hawaiian and USAPI cases for the neuraminidase H275Y mutation associated with oseltamivir resistance by pyrosequencing. RESULTS: Two hundred and sixty-three (263) 2009 pandemic influenza A (H1N1) positive specimens were tested and 263/263 (100%) were shown to lack the mutation most commonly associated with oseltamivir resistance. CONCLUSIONS: There was no evidence of oseltamivir resistant A(H1N1)pdm09 virus during the 2009 pandemic in the Pacific islands despite considerable travel exposure. Geographic isolation, the lack of a "second wave" of pandemic influenza, judicious antiviral use, aggressive vaccination, and below average tourism due to the global economic crisis may have been contributing factors. Continued surveillance and vigilance is necessary to monitor unpredictable influenza activity.

Specimens from a vesicular lesion caused by molluscum contagiosum virus produced a cytopathic effect in cell culture that mimicked that produced by herpes simplex virus.

Infection with molluscum contagiosum virus, a poxvirus, normally has a typical clinical presentation; therefore, laboratory confirmation is infrequently sought and the virus is rarely isolated in culture. As reported herein, viral culture of specimens from atypical lesions may produce an abortive infection in limited cell lines and a cytopathic effect suggestive of herpes simplex virus.