COVID-19 Response Efforts of Washington State Public Health Laboratory: Lessons Learned.

Laboratory diagnostics play an essential role in pandemic preparedness. In January 2020, the first US case of COVID-19 was confirmed in Washington State. At the same time, the Washington State Public Health Laboratory (WA PHL) was in the process of building upon and initiating innovative preparedness activities to strengthen laboratory testing capabilities, operations, and logistics. The response efforts of WA PHL, in conjunction with the Centers for Disease Control and Prevention, to the COVID-19 outbreak in Washington are described herein-from the initial detection of severe acute respiratory syndrome coronavirus 2 through the subsequent 2 months.Factors that contributed to an effective laboratory response are described, including preparing early to establish testing capacity, instituting dynamic workforce solutions, advancing information management systems, refining laboratory operations, and leveraging laboratory partnerships. We also report on the challenges faced, successful steps taken, and lessons learned by WA PHL to respond to COVID-19.The actions taken by WA PHL to mount an effective public health response may be useful for US laboratories as they continue to respond to the COVID-19 pandemic and may help inform current and future laboratory pandemic preparedness activities.

Laboratory Review of Foodborne Disease Investigations in Washington State 2007-2017.

The Washington State Department of Health Public Health Laboratories (WAPHL) has tested 11,501 samples between 2007 and 2017 for a foodborne disease using a combination of identification, serotyping, and subtyping tools. During this period there were 8037 total clinical and environmental samples tested by pulsed-field gel electrophoresis (PFGE), including 512 foodborne disease clusters and 2176 PFGE patterns of Salmonella enterica subsp. enterica. There were 2446 Shiga toxin-producing Escherichia coli samples tested by PFGE, which included 158 foodborne disease clusters and 1174 PFGE patterns. There were 332 samples of Listeria monocytogenes tested by PFGE, including 35 foodborne disease clusters and 104 PFGE patterns. Sources linked to outbreaks included raw chicken, unpasteurized dairy products, various produce types, and undercooked beef among others. As next-generation sequencing (NGS) replaces PFGE, the impact of this transition is expected to be significant given the enhanced cluster detection power NGS brings. The measures presented here will be a reference baseline in future years.

Rapid identification of mycobacteria and drug-resistant Mycobacterium tuberculosis by use of a single multiplex PCR and DNA sequencing.

Tuberculosis (TB) remains a significant global health problem for which rapid diagnosis is critical to both treatment and control. This report describes a multiplex PCR method, the Mycobacterial IDentification and Drug Resistance Screen (MID-DRS) assay, which allows identification of members of the Mycobacterium tuberculosis complex (MTBC) and the simultaneous amplification of targets for sequencing-based drug resistance screening of rifampin-resistant (rifampin(r)), isoniazid(r), and pyrazinamide(r) TB. Additionally, the same multiplex reaction amplifies a specific 16S rRNA gene target for rapid identification of M. avium complex (MAC) and a region of the heat shock protein 65 gene (hsp65) for further DNA sequencing-based confirmation or identification of other mycobacterial species. Comparison of preliminary results generated with MID-DRS versus culture-based methods for a total of 188 bacterial isolates demonstrated MID-DRS sensitivity and specificity as 100% and 96.8% for MTBC identification; 100% and 98.3% for MAC identification; 97.4% and 98.7% for rifampin(r) TB identification; 60.6% and 100% for isoniazid(r) TB identification; and 75.0% and 98.1% for pyrazinamide(r) TB identification. The performance of the MID-DRS was also tested on acid-fast-bacterium (AFB)-positive clinical specimens, resulting in sensitivity and specificity of 100% and 78.6% for detection of MTBC and 100% and 97.8% for detection of MAC. In conclusion, use of the MID-DRS reduces the time necessary for initial identification and drug resistance screening of TB specimens to as little as 2 days. Since all targets needed for completing the assay are included in a single PCR amplification step, assay costs, preparation time, and risks due to user errors are also reduced.

Lake-associated outbreak of Escherichia coli O157:H7 in Clark County, Washington, August 1999.

CONTEXT: Escherichia coli O157:H7, one of hundreds of strains of the gram-negative bacterium E coli, has been implicated in numerous lake-borne outbreaks of infection during the past decade. In August 1999, several children who later became ill with E coli O157:H7 infection reported swimming in a lake in Clark County, Washington. The lake was closed and an investigation begun. OBJECTIVES: To identify the source of the outbreak and determine risk factors for infection with E coli O157:H7.Design, Setting, and Patients Two case-control studies were performed among residents of and visitors to Clark County in August 1999 by using community and campground-registrant control subjects. Main Outcome Measure Risk factors for infection with E coli O157:H7 among Clark County residents or visitors. RESULTS: We identified 37 case patients (including 29 primary-case patients) with a median age of 5 years (age range, 1-14 years for primary-case patients). Eight children were hospitalized, 3 with hemolytic uremic syndrome; none died. With analysis restricted to primary-case patients, illness was strongly associated with swimming in the lake (18 of 18 case patients vs 1 of 18 neighborhood-matched and age-matched control subjects; matched odds ratio undefined; P<.001). All primary-case patients were children younger than 15 years who swam in the lake. Illness was associated with placing the head underwater, getting lake water in the mouth, or swallowing lake water (26 of 27 case patients vs 43 of 62 control subjects; matched odds ratio = 11.5; P =.005). Cultures of lake water yielded E coli O157:H7 that matched the outbreak strain according to results of pulsed-field gel electrophoresis. CONCLUSIONS: To date, this is one of the largest documented outbreaks of E coli O157:H7 infection associated with unchlorinated recreational water and represents the first outbreak in which the strain was isolated from lake water. Guidelines are needed to decrease the risk of enteric illness associated with swimming in recreational lakes.

Obligate bacterial endosymbionts of Acanthamoeba spp. related to the beta-Proteobacteria: proposal of 'Candidatus Procabacter acanthamoebae' gen. nov., sp. nov.

All obligate bacterial endosymbionts of free-living amoebae currently described are affiliated with the alpha-Proteobacteria, the Chlamydiales or the phylum Cytophaga-Flavobacterium-Bacteroides. Here, six rod-shaped gram-negative obligate bacterial endosymbionts of clinical and environmental isolates of Acanthamoeba spp. from the USA and Malaysia are reported. Comparative 16S rDNA sequence analysis demonstrated that these endosymbionts form a novel, monophyletic lineage within the beta-Proteobacteria, showing less than 90% sequence similarity to all other recognized members of this subclass. 23S rDNA sequence analysis of two symbionts confirmed this affiliation and revealed the presence of uncommon putative intervening sequences of 146 bp within helix-25 that shared no sequence homology to any other bacterial rDNA. In addition, the 23S rRNA of these endosymbionts displayed one polymorphism at the target site of oligonucleotide probe BET42a that is conserved in all other sequenced beta-Proteobacteria. Intra-cytoplasmatic localization of the endosymbionts within the amoebal host cells was confirmed by electron microscopy and fluorescence in situ hybridization with a specific 16S rRNA-targeted oligonucleotide probe. Based on these findings, the provisional name 'Candidatus Procabacter acanthamoebae' is proposed for classification of a representative of the six endosymbionts of Acanthamoeba spp. studied in this report. Comparative 18S rDNA sequence analysis of the Acanthamoeba host cells revealed their membership with either Acanthamoeba 18S rDNA sequence type T5 (Acanthamoeba lenticulata) or sequence type T4, which comprises the majority of all Acanthamoeba isolates.