IAI INTRA-ABDOMINAL INFECTION APPLICATIONarrowdiagnostics.it/download/microbiologia...DNA purification PCR set-up Multiplex PCR with array detection 1. Unyvero T1/B1 sample Tube for

Caution – Investigational Device, Limited by Federal (or US) Law to Investigational Use. Not available for Sale in the United States.

IAI INTRA-ABDOMINALINFECTIONAPPLICATION

2 3

CONTeNT


UNYVERO IAI APPLICATION

The Unyvero IAI Application is a high-quality

tool to detect pathogens (bacteria and fungi),

toxins, and antibiotic resistance markers in

severe intra-abdominal infections. The Unyvero

IAI Cartridge analyzes simultaneously in one

patient sample over 100 DNA targets. In a few

hours, specific analytes for resistance markers,

pathogens, toxins and also a Universal Bacteria

primer are assayed. Intra-abdominal infections

are associated with unmet medical needs. The

Unyvero approach helps to make informed

treatment decisions at an earlier stage.

INTRA-ABDOMINAL INFECTIONs 04 – 05

ANTIbIOTIC ReSISTANCeS ARe INCReASINg 06 – 07

UNYVERO IAI INTRA-ABDOMINAL

INFECTION APPLICATION 08 – 09

UNyveRO SySTem – UNIveRSAL PLATFORm 10

COVERAGE – DIVERsITY – RELIABILITY 11

A COmPReheNSIve PANeL 12 – 13

NATIVE sAMPLE TYPEs 14 – 15

eSTAbLISheD TeChNOLOgIeS

IN A New FORmAT 16 – 17

UNYVERO wORkFLOw 18 – 19

UNyveRO SySTem AND APPLICATIONS 20 – 21

UNYVERO AT A GLANCE 22 – 23

4 5Caution – Investigational Device, Limited by Federal (or US) Law to Investigational Use. Not available for Sale in the United States.

INTRA-ABDOMINAL INFECTIONSeCOND mOST COmmON CAUSe OF mORTALITy FROm INFeCTIONS IN INTeNSIve CARe

Improving treatment of patients with

acute abdomen, including peritonitis,

acute pancreatitis and megacolon

requires rapid pathogen identification.

Intra-abdominal infections (IAI), associated with abdominal

cavity exudations, elicit within the peritoneum complex

inflammatory responses to microorganisms and their

toxins. IAI is the second most common type of infections in

patients with severe sepsis and ranks second as cause of

death in US intensive care units.

In addition, complicated IAIs, with a yearly incidence rate

of 300,000 cases in the US, present a significant burden to

health care systems.

INTRA-ABDOMINAL INFECTIONs ARE

CLAssIFIED As:

> community-acquired intra-abdominal infection (CA-IAI)

> healthcare-acquired intra-abdominal infection (hA-IAI)

The acute abdomen, a case of emergency, often caused

by severe infections and necessitate timely medical

attention.

Sources of infections are mostly the appendix, gall

bladder, or are associated with colon or gastro-duodenal

perforations, or diverticulitis.

INTRA-AbDOmINAL INFeCTION

The medical challenge of today is to treat complex IAI

infections that are due to multiple pathogens, together

with the significant risk of complications. Therefore, in

cases of nosocomial intra-abdominal infections, the first

choice of antimicrobial treatment is of major importance.

According to various publications, unsuitable antimicrobial

therapy seems to be linked to longer inpatient residence

and higher fatality rates.

An increasing resistance to gram–negative pathogens,

which are responsible for serious nosocomial infections,

e.g. intra-abdominal infections such as:

> ß-lactam resistance Enterobacteriacea

> carbapenem resistance Enterobacteriacea

> mDR P. aeruginosa

is the result of an increasing acquisition of resistance

genes worldwide.

The global alliance to optimize rational use of antimicrobials

for intra-abdominal infections (AgORA) recommended:

An optimal antimicrobial approach to treating IAI involves

a delicate balance between the optimization of empiric

therapy, which improves clinical outcomes, and the

reduction of excessive antimicrobial use that increases the

rate of emergence of antimicrobial-resistant strains.

early pathogen and antibiotic resistance gene identification

with rapid molecular tests may overcome those problems.


ANTIBIOTIC REsIsTANCEgeT The beTTeR OF A SeRIOUS PRObLem by eARLy IDeNTIFICATION

ANTIbIOTIC ReSISTANCeS ARe INCReASINg

DNA purification

PCR set-up

Multiplex PCRwith array detection

1. Unyvero T1/B1 sample Tube for preparation of the patient sample, pre-filled with specific lysis reagents.

2. Unyvero T1/B1 sample Tube Cap seals the Unyvero Sample Tube and contains proteinase K and an internal control gene for quality control.

3. Unyvero T1 sample Pre-Treatment Tool is an aid for preparation and processing of sample materials.

4. Unyvero M1 Master Mix Tube containing reagents for DNA amplification.

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THE UNIQUE UNYVERO IAI CARTRIDGE COVERs

PATHOGENs, ANTIBIOTIC REsIsTANCE MARkERs

AND TOXINs, AND COMBINEs

> DNA purification

> PCR set-up

> multiplex endpoint PCR

> detection by array hybridization

Therefore, the closed Unyvero Cartridge is equipped

with integrated reagent containers, a DNA purification

column, eight PCR chambers, and an according number

of detection arrays.

The cartridge is pre-filled with buffers for DNA clean-up,

reagents, and fluorescence-labeled primers for PCR ampli-

fication, and for array hybridization of the PCR product.3

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sOLUTION – UNYVERO IAIAPPLICATION

UNIQUE sOLUTION

TAILORED TO CLINICAL NEEDs

> Comprehensive, disease-specific test panels

run on an easy to operate platform, requiring

neither the special infrastructure of a laboratory

environment nor specially trained personnel.

> Translates a complex laboratory process into

an easy to use format. In addition to the un-

processed patient sample, only enzymes for

DNA amplification are required for the analytical

process.

> Generates complete diagnostic information

in 4 – 5 hours, without any further operator

interaction.

UNyveRO IAI INTRA-AbDOmINAL INFeCTION APPLICATION

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UNyveRO SySTem AND APPLICATIONS

INTRA-ABDOMINAL INFECTION AssAY

> DETECTION OF COMPLICATED

INTRA-ABDOMINAL INFECTIONs

> ANY INFECTION FOCUs

> VARIOUs NATIVE sAMPLE TYPEs

> OVER 100 TARGETs

> 1 CARTRIDGE

3. sPEED

Delivers clinically relevant

answers in time

4 – 5 H

2. DIVERsITY

Enables testing of clinically

relevant sample types of

abdominal cavity

> BILE, PERITONEAL

FLUID AND AsCITEs

> TIssUE

> swABs, CATHETER

TIPs

> POsITIVE IAI FLUID

IN BLOOD CULTURE

BOTTLE

1. COVERAGE

A selection of pathogens,

toxins, and antibiotic

resistances based

on incidence, clinical

importance, and treatment

guidlines

> OVER 100 TARGETs

PReReqUISITeS


UNYVERO sYsTEM – UNIVERsAL PLATFORMFeATURINg mANy DIFFeReNT APPLICATIONS

UNYVERO APPLICATION CARTRIDGEs

UNYVERO A50 ANALYzERUNYVERO C8 COCkPITUNYVERO L4 LYsATOR

GOLDEN HOUR

In the face of emerging multiresistant organisms, antimicrobioal

selection becomes more and more difficult, because many

different genetic mechanisms are involved. Therefore, testing a

broad spectrum with over 100 targets is required to get accurate

information for guiding adequate, targted therapy.

UNYVERO HELPs TO

> focus on the appropriate therapy

> choose the right antibiotic

> improve disease outcome

1 incl. S. saprophyticus, S. hominis, S. epidermidis, S. warneri, S. haemolyticus, S. capitis, S. lugdunensis

2 incl. E. faecalis, E. faecium, E. gallinarum, E. casseliflavus, E. avium, E. hirae, E. durans, E. raffinosus

3 incl. S. pneumoniae, S. constellatus, S. agalactiae, S. pyogenes, S. anginosus4 incl. A. hydrophila subsp. hydrophila, A. sobria, A. media, A. veronii, A. caviae5 incl. B. fragilis, B. ovatus, B. thetaiotaomicron, B. uniformis6 incl. B. fragilis, B. ovatus, B. heparinolyticus, B. pyogenes, B. thetaiotaomicron,

B. uniformis, B. vulgatus, Parabacteroides distasonis, P. bivia, P. buccae, P. disiens, P. intermedia, P. jejuni, P. melaninogenica, P. nigrescens, P. oris, P. denticola

7 incl. E. cloacae, E. asburiae, E. hormaechei8 incl. Klebsiella pneumoniae Cluster kp I + II, Alves et al., J Clin Microbiol, 44(10),

2006; 9 incl. Klebsiella variicola (Cluster kpIII; formerly K. pneumoniae, 2004 classified as its

own species10 incl. P. vulgaris, P. mirabilis, P. penneri, P. hauseri11 incl. A. baumannii, A. oleivorans, A. calcoaceticus, A. pittii

GROUP PATHOGEN

Universal bacteria

gram-positive bacteria

Coagulase negative staphylococci (CNS) 1

Enterococcus faecalis

Enterococcus spp. 2

Streptococcus spp. 3

Staphylococcus aureus

Anaerobic / facultative anaerobic bacteria

Aeromonas spp. 4

Bacteroides fragilis group 5

Bacteroides spp. / Prevotella spp. 6

Clostridium difficile

Clostridium perfringens

Finegoldia magna

Propionibacterium acnes

Enterobacteriaceae

Escherichia coli

Enterobacter cloacae complex 7

Klebsiella pneumoniae 8

Klebsiella oxytoca

Klebsiella variicola 9

Proteus spp. 10

GROUP PATHOGEN

Non-fermenting bacteriaAcinetobacter baumannii complex 11

Pseudomonas aeruginosa

Fungi

Candida spp.

Candida albicans

Candida glabrata

Candida tropicalis

Issatchenkia orientalis (Candida krusei)

GENE REsIsTANCE AGAINsT

mecA Oxacillin

mecC Oxacillin

vanA vancomycin

vanB vancomycin

aacA4 Aminoglycoside

ctx-m 3rd generation Cephalosporin

fosA3 Fosfomycin

mcr-1 Polypeptides/polymyxins

nimA Nitroimidazole

nimb Nitroimidazole

qnrA Fluoroquinolone

qnrb Fluoroquinolone

qnrS Fluoroquinolone

tetA Tetracycline

kpc Carbapenem

oxa-23 Carbapenem

oxa-24/40 Carbapenem

oxa-48 Carbapenem

oxa-58 Carbapenem

vim Carbapenem

imp Carbapenem

ndm Carbapenem

TOXIN MARkER TOXIN

tcdb Toxin b

stx1/2 Shiga Toxin

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A COmPReheNSIve PANeL

COVERAGE - ANTIBIOTIC REsIsTANCEs, CLINICALLY RELEVANT PATHOGENs AND TOXINsINCLUDINg UNIveRSAL bACTeRIA PRImeR

THE COMPREHENsIVE UNYVERO INTRA-

ABDOMINAL INFECTION PANEL COVERs

MANY CLINICALLY RELEVANT PATHOGENs –

UNIVERsAL BACTERIA AND Candida sPP. –

AND IMPORTANT ANTIBIOTIC REsIsTANCE

MARkERs AND TOXINs



wIDE RANGE OF NATIVE sAMPLE TYPEs OveRCOmINg ChALLeNgeS IN DIAgNOSINg IAI

Intra-abdominal infections include a wide range of clinical

pictures ranging from infections of the peritoneum,

pancreas and colon, requiring the testing of many different

and complex sample types, of various natures and

qualities.

The Unyvero IAI application improves the identification

of pathogens and resistance genes in acute bacterial

infections of the abdomen, and uniquely combines it with

the detection of toxin markers in one test. It facilitates

testing of ascites- and peritoneal fluid, pancreatic juice, as

well as bile. Additionally, it handles e.g. catheter tips and

drainage fluid, swabs tissue, and samples from positive

flagged blood cultures.

Compared to traditional IAI diagnostic measures, which

are laborious and time consuming based on growing

cultures and microscopy, multiple closely related non-

pathogenic organisms, prior medication, and the number

of involved pathogens no longer pose a problem.

NATIve SymPLe TyPeS

BILE

PERITONEAL

FLUID

sAMPLEs

FROM BLOOD

CULTURE

BOTTLEs

DRAINAGE

FLUID

swABs

TIssUE

BIOPsY

AsCITEs

BECAUsE OF ITs UNIVERsAL APPLICABILITY

THE CURETIs TECHNOLOGY CAN UsE A sINGLE

sAMPLE PREPARATION PROTOCOL FOR MANY

DIFFERENT sAMPLE TYPEs OF ANY CLINICAL

APPLICATION

One of Curetis‘ core competencies is the design of reliable

bio-assays in self-contained cartridges to enable sensitive

multiplex testing. The required multiplex grade is achieved

by combining the unmatched sample preparation and PCR

technology with a proprietary detection array. endpoint-

PCR can deliver higher multiplex grades than most other

amplification methods, and in that respect holds clear

advantages over e.g. real-time PCR.

The array was optimized for hybridization times to

only a few minutes compared to standard arrays. To

enhance specificity, the dissociation kinetics of the array

hybridization are automatically assessed. Additionally with

each patient sample, an artificial gene is simultaneously

processed as internal process control. The control verifies

all PCR steps, as well as the array hybridization.

> liquid> samples cultured in blood culture bottles> tissue ...

sample Types

Pathogen DNAspecific

Amplificationspecific Target Hybridization

Detection and specific Control

toxin B

Candida albicans

Clostridium difficile

sample Quality

Curetis‘ proprietary and universal sample preparation

technology prepares DNA from any native clinical sample

type without losing time needed for preculturing.

16 17

DIVERsITY – HIGHLY sPECIFIC MULTIPLEX PCRAND FAST ARRAy-bASeD DeTeCTION


eSTAbLISheD TeChNOLOgIeS IN A New FORmAT

The Unyvero IAI Application combines all analytical

steps in one closed test format, thus reducing the work

involved for testing. Only a few simple manual steps allow

simultaneous detection of various microorganisms, toxins,

and antibioticresistance markers simultaneously in native

patient samples and samples from positive flagged blood

cultures within approximately five hours.

The intuitive simple workflow is consistent across all clinical applications and

sample types, thus drastically reducing invalid tests due to operator errors.

sTEP 1 TRANSFeR OF SAmPLeS AND LySIS

sTEP 2 ASSembLINg The UNyveRO CARTRIDge

sTEP 4 FOUR TO FIve hOURS ANALySIS PROCeSS

sTEP 3 SCANNINg AND INSeRTINg The UNyveRO CARTRIDge

~ 30 seconds:

2. After 30 minutes the Unyvero Sample Tube is removed from the Lysator. Then the IAI Cartridge is assembled with the Unyvero Sample Tube and Unyvero master mix Tube.

~ 20 seconds:

3. The Cartride is scanned and insert into the Unyvero Analyzer. Two cartridges per analyzer can be analyzed simultaneously.

It takes just a few minutes for subsequent result analysis:

4. The Unyvero analysis runs for four to five hours. The results are displayed on the screen or can be exported as a PDF.

~ 60 seconds:

1. The patient sample is transferred to the Unyvero Sample Tube, sealed in with the Unyvero Sample Tube Cap, and lysed for 30 minutes in the Unyvero Lysator.

working time

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sPEED – UNYVERO sOLUTIONSImPLIFIeD wORKFLOw

UNyveRO wORKFLOw UNyveRO wORKFLOw


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UNYVERO sYsTEM – UNIVERsAL PLATFORMFeATURINg mANy DIFFeReNT APPLICATIONS

UNyveRO SySTem AND APPLICATIONS

THE UNYVERO sOLUTION FEATUREs A

»sAMPLE TO ANswER« APPROACH AND

CONsIsTs OF:

> the Unyvero L4 Lysator for sample lysis

> the application specific Unyvero Cartridge

> the Unyvero A50 Analyzer prozessing the

Unyvero Cartridge

> the Unyvero C8 Cockpit for intuitive user

interaction

The Unyvero System translates a complex laboratory

process into an easy-to-use format. The analytical

process requires merely the addition of the unprocessed

patient sample and enzymes for DNA amplification.

All other reagents are preloaded into a self-contained,

contamination-safe cartridge. After the patient sample

and the master mix are inserted into the cartridge, the

cartridge is loaded into the analyzer and processed fully

automatically. The Unyvero System generates complete

diagnostic information in 4 – 5 hours, without any further

operator interaction.


UNYVEROAT A gLANCe

THE UNYVERO sOLUTION

> detects a broad panel of pathogens and antibiotic

resistances, as well as toxins from a single sample in

one run

> enables the testing of many clinically relevant native

samples

> delivers clinically relevant information in 4 – 5 hours,

helps with fact-based treatment decisions as early

as possible

> allows point of need testing

> provides high productivity by

- minimizing operator time to a few minutes with

full walk-away automation, based on a simple and

consistent workflow for any clinical application

- low implementation requirements and low total

ownership cost, as neither skilled molecular biology

staff nor special infrastructure is needed

- configurations that meet individual customer needs,

satisfying processing time requirements

- optimizing routine hospital workflow by sophisticated

information management

Caution – Investigational Device, Limited by Federal (or US) Law to Investigational Use. Not available for Sale in the United States.22 23

AT A gLANCe

Curetis gmbh | max-eyth-Str. 42 | 71088 holzgerlingen | germany

Tel.: +49 (0)7031 49195 10 | email: [email protected]

www.unyvero.com | www.curetis.com

0041

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0Caution – Investigational Device, Limited by Federal (or US) Law to Investigational Use. Not available for Sale in the United States.

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IAI INTRA-ABDOMINAL INFECTION APPLICATIONarrowdiagnostics.it/download/microbiologia...DNA purification PCR set-up Multiplex PCR with array detection 1. Unyvero T1/B1 sample Tube for