VTEC-RPLA SEIKEN

K981734 · Denka Seiken'S · GNA · Sep 22, 1998 · Microbiology

Device Facts

• Record ID: K981734
• Device Name: VTEC-RPLA SEIKEN
• Applicant: Denka Seiken'S
• Product Code: GNA · Microbiology
• Decision Date: Sep 22, 1998
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 866.3255
• Device Class: Class 1

Intended Use

This in vitro diagnostic procedure is intended for the detection and identification of verotoxins or shiga-like toxins in culture isolates of E. coli. Such characterization is useful in the diagnosis of Enterohemorrhagic E. coli (EHEC) infections.

Device Story

VTEC-RPLA "SEIKEN" is an in vitro diagnostic reagent system for identifying verotoxins (VT1 and VT2) in E. coli culture isolates. The device utilizes reversed passive latex agglutination (RPLA) technology; latex particles are sensitized with polyclonal antibodies specific to VT1 and VT2. When culture supernatant containing verotoxins is mixed with the sensitized latex, agglutination occurs, indicating a positive result. The test is performed in a laboratory setting by trained personnel. Results are interpreted visually by the clinician to aid in diagnosing EHEC infections. The device allows for specific identification of toxin types, assisting in clinical decision-making regarding patient management and infection control.

Clinical Evidence

Clinical performance established via two studies. Study 1 (in-house): 100 E. coli strains (80 verotoxin-producing, 20 non-producing) tested in parallel with Premier EHEC; 100% sensitivity and 100% specificity observed. Study 2 (external): 178 E. coli strains (147 verotoxin-producing, 31 non-producing) tested using VTEC-RPLA, PCR, and vero cell assay; 100% sensitivity and 100% specificity observed. Bench testing confirmed reproducibility, stability (1-year shelf life at 2-10°C), and consistent agglutination titers (1:64 to 1:128) across reagent lots and operators.

Technological Characteristics

Reagent system based on reversed passive latex agglutination (RPLA). Utilizes polyclonal antibodies specific for VT1 and VT2. Minimal detectable concentration is approximately 25 pg/well. Requires CA-YE shaking broth for specimen preparation. Manual visual interpretation of agglutination titers. No electronic components or software.

Indications for Use

Indicated for the detection and identification of verotoxins in culture isolates of E. coli to aid in the diagnosis of Enterohemorrhagic E. coli (EHEC) infections. Intended for prescription use.

Regulatory Classification

Identification

Escherichia coli serological reagents are devices that consist of antigens and antisera used in serological tests to identify Escherichia coli from cultured isolates derived from clinical specimens. Additionally, some of these reagents consist of Escherichia coli antisera conjugated with a fluorescent dye used to identify Escherichia coli directly from clinical specimens or cultured isolates derived from clinical specimens. The identification aids in the diagnosis of diseases caused by this bacterium belonging to the genus Escherichia, and provides epidemiological information on diseases caused by this microorganism. Although Escherichia coli constitutes the greater part of the microorganisms found in the intestinal tract in humans and is usually nonpathogenic, those strains which are pathogenic may cause urinary tract infections or epidemic diarrheal disease, especially in children.

Predicate Devices

• Premier EHEC (608096)

Reference Devices

• PCR (not specified)
• vero cell assay (not specified)

Submission Summary (Full Text)

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SEP 2 2 1998

. ... ..

K981734

## ATTACHMENT B

# SUMMARY OF SAFETY AND EFFECTIVENESS

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## SUMMARY OF SAFETY AND EFFECTIVENESS

## VTEC-RPLA "SEIKEN"

Below summarizes and compares the performance of VTEC-RPLA "SEIKEN" and a similar device previously given FDA clearance for marketing in the US. The information contained in this summary was obtained from data prepared at and which is on file at Denka Seiken Co., Inc. This summary shows that the two reagent systems are substantially equivalent.

## INTENDED USE

This in vitro diagnostic procedure is intended for the detection and identification of verotoxins or shiga-like toxins in culture isolates of E. coli. Such characterization is useful in the diagnosis of Enterohemorrhagic E. coli (EHEC) infections.

| METHOD | VTEC-RPLA "SEIKEN" | Premier EHEC |
|-----------------------|--------------------|--------------|
| Product code | 230553 | 608096 |
| Min. Detectable Conc. | | |
| VT 1 | 25 pg/well | 7 pg/well |
| VT2 | 25 pg/well | 15 pg/well |

Correlation

Specificity and sensitivity between the two tests were 100% and 100%, respectively.

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## VTEC-RPLA "SEIKEN" and Premier EHEC are similar in that both:

- * Are reagent systems which can be used for the detection of verotoxins in culture isolates of E. coli to aid in the diagnosis of EHEC infections.
- * Employ antibody-antigen reactions as part of the assay.

#### VTEC-RPLA "SEIKEN" and Premier EHEC are different in that:

- * They use different culture conditions to prepare specimens
- * VTEC-RPLA is based on reversed passive latex agglutination (RPLA) while the Premier EHEC is an enzyme immuno assay (EIA).
- * The minimal detectable concentration for the VTEC-RPLA is approximately 25 pg / well ; that for the Premier EHEC is 7- 15 pg / well.
- * VTEC-RPLA utilizes polyclonal antibodies specific for each verotoxin type, VT1 and VT2, and thus may be used to identify toxins; the Premier EHEC uses three types of antibody: an anti-VT monoclonal (EIA capture); anti-VT polyclonal (enzyme conjugate); and anti-rabbit IgG goat IgG conjugated with POD; the assay is specific for verotoxins but not type specific, ie cannot identify VT1 and VT2.
- * Premier EHEC may be used to directly detect verotoxins in stools, in mixed cultures and in culture isolates while VTEC-RPLA is intended for use with culture isolates only.

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## PROTOCOL AND DATA SUMMARY

This section provides data generated by and for Denka Seiken Co., Ltd. characterizing the performance of VTEC-RPLA "SEIKEN". The protocols used for the data generation are given below, and the results are attached.

## CORRELATION

#### Study 1 (in-house)

One hundred Eschericia coli strains (80 verotoxin-producing and 20 non-verotoxin producing) were obtained from the Japanese National Institute of Infectious Disease and tested in parallel using VTEC-RPLA "SEIKEN" and the Premier EHEC Reagent System. Specimens were prepared and tested according to the product inserts given in the kits. 100% sensitivity and 100% specificity were observed. Study 2 (external)

One hundred and seventy eight Eschericia coli strains (147 verotoxin-producing and 31 non-verotoxin producing) isolated from humans were tested by the Tokyo Metropolitan Research Laboratory of Public Health using VTEC-RPLA "SEIKEN", PCR and the vero cell assay. 100% sensitivity and 100% specificity were observed.

## SENSITIVITY

Three reagent lots (Lot No. 3421, No. 3626 and No. 31222) and two verotoxinproducing strains, DK-EC-VT1 and DK-EC-VT2, obtained from Kyoto University Faculty of Medicine, were used in these studies. Verotoxin type 1 and verotoxin type 2 were purified from the respective strains above and preparations were adjusted to 100 ng/ml for testing. Each verotoxin preparation was tested with each reagent lot three times and all three lots showed titers of between 1:64 to 1:128 in all replicates.

## SPECIFICITY

Three reagent lots (Lot No. 3421, No. 3626 and No. 31222) , eight verotoxin-producing and twelve non-producing strains of E. coli were used in these studies. Specimens were prepared using the CA-YE shaking broth recommended in the product insert and the resulting culture supernatants from each strain were lyophilized and subsequently tested in triplicate after reconstitution. Positive results were obtained from verotoxin-producing strains and, likewise, negative results only from non-producing strains.

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#### REPRODUCIBILITY

Reagent lots and verotoxin specimens were the same as those described in SEN-SITIVITY above. Each verotoxin preparation was tested with each reagent lot five times and the resulting titers for all assays were between 1:64 to 1:128. Also, performed were Person--to--person: testing done by three operators Day--to--day: testing done on five different days Lot -- to -- lot: testing done with three different lots All of the above studies showed agglutination titers of between 1:64 to 1:128.

#### DILUTION

Three verotoxin-producing strains, DK-EC-PS 1 (VT1), DK-EC- PS 4 (VT2) and DK-EC-PS 7 (VT1 and VT2), were cultured in CA-YE by the broth culture method and the supernatants obtained after cell-pelleting, in addition to ten-fold dilutions in saline, were each tested in a single examination. All ten-fold dilutions showed an agglutination titer at the expected range.

#### STABILITY

To establish storage conditions and the reagent shelf-life, the above three reagent lots were tested at 0, 3, 6, 9, 12 and 15 months with regards to sensitivity, specificity and reproducibility as described above. Reagents were store at 12 C. As there was no change in reagent performance throughout this time, the storage conditions and shelf-life were set at 2-10 C and one year after production, respectively.

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Image /page/5/Picture/2 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo features a stylized eagle with its wings spread, and the words "DEPARTMENT OF HEALTH & HUMAN SERVICES USA" are arranged in a circular pattern around the eagle. The logo is black and white and appears to be a scanned image.

## SEP 2 2 1998

Food and Drug Administration 2098 Gaither Road Rockville MD 20850

Kevin Mangan International Sales and Business Development Denka Seiken Co., Ltd. 3-4-2. Nihonbashi-Kavabacho Chuo-Ku Tokyo, Japan 103-0025

Re: K981734 Trade Name: VTEC-RPLA "SEIKEN" Regulatory Class: I Product Code: GNA Dated: August 14, 1998 Received: August 17, 1998

Dear Mr. Mangan:

We have reviewed your Section 510(k) notification of intent to market the device referenced above and we have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act). You may, therefore, market the device, subject to the general controls provisions of the Act. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration.

If your device is classified (see above) into either class II (Special Controls) or class III (Premarket Approval), it may be subject to such additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations. Title 21, Parts 800 to 895. A substantially equivalent determination assumes compliance with the Current Good Manufacturing Practice requirements, as set forth in the Quality System Regulation (QS) for Medical Devices: General regulation (21 CFR Part 820) and that, through periodic QS inspections, the Food and Drug Administration (FDA) will verify such assumptions. Failure to comply with the GMP regulation may result in regulatory action. In addition, FDA may publish further announcements concerning your device in the Federal Register. Please note: this response to your premarket notification submission does not affect any obligation you might have under sections 531 through 542 of the Act for devices under the Electronic Product Radiation Control provisions, or other Federal laws or regulations.

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#### Page 2

Under the Clinical Laboratory Improvement Amendments of 1988 (CLIA-88), this device may require a CLIA complexity categorization. To determine if it does, you should contact the Centers for Disease Control and Prevention (CDC) at [phone].

This letter will allow you to begin marketing your device as described in your 510(k) premarket notification. The FDA finding of substantial equivalence of your device to a legally marketed predicate device results in a classification for your device and thus, permits your device to proceed to the market.

If you desire specific advice for your device on our labeling regulation (21 CFR Part 801 and additionally 809.10 for in vitro diagnostic devices), please contact the Office of Compliance at [phone]. Additionally, for questions on the promotion and advertising of your device, please contact the Office of Compliance at [phone]. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR 807.97). Other general information on your responsibilities under the Act may be obtained from the Division of Small Manufacturers Assistance at its toll free number [phone] or at [phone] or at its internet address "http://www.fda.gov/cdrh/dsmamain.html"

Sincerely yours,

Steven Autman

Steven I. Gutman, M.D., M.B.A. Director Division of Clinical Laboratory Devices Office of Device Evaluation Center for Devices and Radiological Health

Enclosure

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Page -

510(k) Number (ii known) K981734

Device Name: VTEC-RPLA

Indications For Use

VTEC-RPLA "SEIKEN" is an in vitro diagnostic device for the detection and identification of verotoxic device for in culture isolates of E. coli and is intended as an aid in the diagnosis of Enterohemorrhagic E. coli (EHEC) infections.

FDA/CDRH/ODE/DMC

12 JUN 93
1305

INITIALED

(PLEA
NEEDED

(PLEASE DO NOT WRITE BELOW THIS LINE-CONTINUE ON ANOTHER PAGE IF
EEDED) NEEDED)

Concurrence of CDRH, Office of Device Evaluation (ODE)

Woody Dubois

n of Clinical Laboratory Devices Number

Prescription Use (Per 21 CFR 801.109)

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Gver-The-Counter Use__________________________________________________________________________________________________________________________________________________________

(Optional Format 1-2-96)

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