Lab Matters Summer 2018 | Page 43

APHL 2018 Annual Meeting Poster Abstracts

Whole-Genome Sequencing of Carbapenemase-producing Klebsiella pneumoniae Isolates Recovered from a CRE Colonization Investigation

E . Snavely 1 , E . Nazarian 1 , D . Baker 1 , J . Bodnar 1 , K . Cummings 1 , P . Lapierre 1 , K . Mitchell 1 , S . Morris 1 , J . Shea 1 , C . Wagner 1 , D . Wroblewski 1 , C . B . Kinsey 2 , L . Dettinger 2 , D . Xia 2 , K . Musser 1 ;

1

Wadsworth Center , New York State Department of Health , Albany , NY , 2 Pennsylvania Department of Health , Harrisburg , PA

In this study , whole-genome sequencing ( WGS ) was used to retrospectively analyze carbapenemase-producing carbapenemresistant Enterobacteriaceae ( CP-CRE ) isolates . These isolates were recovered during a point prevalence survey at a 120-bed long term care facility ( LTCF ) in Pennsylvania , where 8 cases of CP-CRE were confirmed at the Centers for Disease Control as

bla KPC

-positive Klebsiella pneumoniae . As the Northeast Regional Antimicrobial Resistance Laboratory Network ( ARLN ) laboratory , the Wadsworth Center is funded to perform CRE colonization testing in response to the detection of CP-CRE . Multiplex real-time PCR assays performed in our laboratory detected the bla KPC gene in 13 of 77 dual rectal swab specimens collected from residents of the PA facility ; 12 unique K . pneumoniae isolates were recovered using colonization culture methods from 10 of the original 13 specimens . The retrospective analysis of these isolates by WGS determined all isolates to be sequence type 258 ( ST258 ) by multilocus sequence typing analysis . WGS analysis also determined that all isolates were positive for the bla KPC

-3 variant , which is associated with a plasmid that harbors other antimicrobial resistance determinants , including genes that confer aminoglycoside , sulfonamide and trimethoprim resistance . Other beta-lactamase families identified during this analysis include OXA ( 83 %), TEM ( 100 %) and SHV ( 100 %). Antimicrobial susceptibility testing by broth microdilution indicated 11 of 12 isolates recovered were resistant to ertapenem , imipenem and meropenem . These 11 isolates were also positive when tested for phenotypic carbapenemase production using the modified carbapenem inactivation method ( mCIM ). WGS investigations of the single mCIM negative , carbapenem-susceptible isolate revealed a nonsense mutation in the bla KPC gene that introduced a premature stop codon . The majority of isolates in this study ( 10 of 12 ) were also resistant to trimethoprim / sulfamethoxazole ( TMP / SMX ). In contrast , 2 of the 12 isolates were sensitive to all aminoglycoside antibiotics , along with TMP / SMX . The susceptibility profile differences of these two isolates correspond with the absence of 4 genes that confer aminoglycoside resistance and 3 genes known to be involved in trimethoprim and sulfonamide resistance ; these genes are present in the WGS data of the other 10 isolates . PFGE analysis revealed 2 clusters formed by 9 isolates , with the remaining 3 of 12 isolates unrelated . Interestingly , WGS SNP analysis identified 1 unrelated isolate and characterized 11 isolates into 2 clusters ; one cluster contained 8 isolates , while the other cluster is formed by 3 isolates . Continued work on this study and in similar studies will contribute to the understanding of the transmission of emerging CP-CRE strains and impact infection control activities .

Presenter : Emily Snavely , PhD , Wadsworth Center , New York State Department of Health , Albany , NY , Phone : 605.431.5044 , Email : emily . snavely @ health . ny . gov

Validation of a Real-time PCR Assay for Rapid Detection of Emerging Multidrug-resistant C . auris from Blood

A . Ojebode , L . Leach and S . Chaturvedi ; Wadsworth Center , New York State Department of Health , Albany , NY

Candida auris is an emerging multidrug-resistant yeast , causing invasive healthcare-associated infections with high mortality in several countries including the US . Mortality associated with C . auris infections are found to be common among the elderly , severely ill patients with indwelling catheters and hematological malignancies . Conventional blood culture considered as “ gold standard ” for candidemia identification , is time-consuming and takes at least 5-7 days . In certain instances , C . auris has been misidentified as C . haemulonii or C . sake by this method as well , thereby making it easy for the oblivious spread of this yeast . To overcome these problems , we utilized our recently developed real-time PCR assay in the laboratory for the validation of blood as a matrix for the detection of C . auris . The limit of detection of the assay was five C . auris CFU / PCR reaction with an efficiency rate of 87 % to 91 %. The assay was highly reproducible as it produced consistent results within and on different days of testing . Overall , our assay is rapid with a turn-around time of six hours as compared to 5 – 7 days with blood culture and present a better approach for managing patients with C . auris induced candidemia as well as the implementation of public health measures to control the spread of this emerging multidrugresistant yeast pathogen .

Presenter : Ayodele Ojebode , MPH , MT ( ASCP ), Wadsworth Center , New York State Department of Health , Albany , NY , Phone : 518.474.2175 , Email : ayodele . ojebode @ health . ny . gov

Reduced Viability of Candida auris and Other Candida Species from MALDI-TOF Extractions

( complete abstract in Infectious Disease , p . 78 )

APHL General

The 2017 APHL Membership Survey : Results Are In !

L . Granen , D . Gaskins and K . Albrecht , Association of Public Health Laboratories , Silver Spring , MD

Introduction : Every 2 -3 years , APHL administers a member satisfaction survey seeking responses and feedback from all APHL members . In 2017 , the membership survey was administered between the dates of January 25 , 2017 and February 23 , 2017 .

Methods : Survey was sent to all APHL members . The Membership Department provided the content for the questions and added new types of questions namely on assessing the real value of APHL membership and member engagement both now and in the future . Survey questions covered members ’ satisfaction levels with a variety of APHL products and services . Survey was administered by APHL ’ s Institutional Research Program .

Results : Response rate for the survey was 31 % ( 265 ). Of the 265 who responded , the highest numbers of respondents were in the state institutional member category ( 51.3 %). Satisfaction with APHL membership was very high , with > 99 % of respondents saying they were satisfied with their benefits . Likewise , the satisfaction rating for interactions with APHL staff was 97.3 % ( excellent or

Antimicrobial Resistance / APHL General