MODIFICATION TO JBAIDS ANTHRAX DETECTION SYSTEM

K071188 · Idaho Technology, Inc. · NHT · May 21, 2007 · Microbiology

Device Facts

• Record ID: K071188
• Device Name: MODIFICATION TO JBAIDS ANTHRAX DETECTION SYSTEM
• Applicant: Idaho Technology, Inc.
• Product Code: NHT · Microbiology
• Decision Date: May 21, 2007
• Decision: SESE
• Submission Type: Special
• Regulation: 21 CFR 866.3045
• Device Class: Class 2

Indications for Use

The JBAIDS Anthrax Detection System is a real-time polymerase chain reaction (PCR) test system intended for the qualitative in vitro diagnostic (IVD) detection of target DNA sequences on the pXO1 plasmid (Target 1) and the pXO2 plasmid (Target 2) from Bacillus anthracis. The system can be used to test human whole blood collected in sodium citrate from individuals suspected of having anthrax, positive blood cultures, and cultured organisms grown on blood agar plates. The JBAIDS Anthrax Target 2 assay is used as a supplementary test only after a positive result with the Target 1 Assay. The JBAIDS Anthrax Target 1 and Target 2 Assays are run on the JBAIDS instrument using the Diagnostic Wizard. Results are for the presumptive identification of B. anthracis, in conjunction with culture and other laboratory tests. The following considerations also apply: The diagnosis of anthrax infection must be made based on history, signs, symptoms, exposure likelihood, and other laboratory evidence, in addition to the identification of pXO1 and pXO2 targets either from cultures or from direct blood specimens. The assays have not been evaluated with blood from individuals without clinical signs or symptoms who were presumed exposed and who subsequently developed anthrax (inhalation or other forms of the disease), or from individuals with any form of anthrax (inhalational, cutaneous, or gastrointestinal). The level of plasmid targets that would be present in blood from individuals with early systemic infection is unknown. The definitive identification of B. anthracis from colony growth, liquid blood culture growth, or from blood specimens requires additional testing and confirmation procedures in consultation with public health or other authorities for whom reports are required. The safety and effectiveness of other types of tests or sample types (not identified as "For in vitro diagnostic use") have not been established.

Device Story

JBAIDS is a portable, integrated IVD system for real-time PCR detection of B. anthracis. Inputs: whole blood, blood culture, or colony samples. Process: sample purification (IT 1-2-3 kits), followed by PCR amplification in glass capillaries using a thermocycler and fluorimeter. Hydrolysis probes detect specific DNA targets (pXO1/pXO2); Taq polymerase hydrolyzes probes upon hybridization, releasing reporter dye. Instrument monitors fluorescence across three wavelengths. Software analyzes amplification curves to report results as Positive, Negative, Inhibited, or Uncertain. Used in clinical laboratory settings by trained personnel. Output aids clinicians in presumptive diagnosis of systemic anthrax when combined with culture and clinical history. Benefits: rapid, automated molecular identification of pathogenic DNA sequences.

Clinical Evidence

Bench testing only. Method comparison study using 6 spiked citrated whole blood samples at LOD showed 100% agreement between new and predicate purification kits. Study of 16 healthy donor samples showed 100% negative results for both methods. Carry-over study (n=42) showed 2.4% carry-over rate for new kit, deemed equivalent to predicate performance.

Technological Characteristics

Real-time PCR system. Modification involves QIAamp spin column-based DNA purification. Reagents include protease K, carrier RNA, and Qiagen buffers (AL, AW1, AW2, AE). System includes JBAIDS instrument, laptop, and software. Operates via benchtop centrifuge for sample prep. Qualitative detection of pXO1/pXO2 plasmids.

Indications for Use

Indicated for qualitative in vitro diagnostic detection of B. anthracis DNA (pXO1 and pXO2 plasmids) in human whole blood (sodium citrate), positive blood cultures, and cultured organisms from blood agar plates in individuals suspected of having anthrax.

Regulatory Classification

Identification

An in vitro diagnostic device for Bacillus species (spp.) detection is a prescription device used to detect and differentiate among Bacillus spp. and presumptively identify B. anthracis and other Bacillus spp. from cultured isolates or clinical specimens as an aid in the diagnosis of anthrax and other diseases caused by Bacillus spp. This device may consist of Bacillus spp. antisera conjugated with a fluorescent dye (immunofluorescent reagents) used to presumptively identify bacillus-like organisms in clinical specimens; bacteriophage used for differentiating B. anthracis from other Bacillus spp. based on susceptibility to lysis by the phage; or antigens used to identify antibodies to B. anthracis (anti-toxin and anti-capsular) in serum. Bacillus infections include anthrax (cutaneous, inhalational, or gastrointestinal) caused by B. anthracis, and gastrointestinal disease and non-gastrointestinal infections caused by B. cereus.

Special Controls

*Classification.* Class II (special controls). The special controls are set forth in FDA's special controls guideline document entitled “In Vitro Diagnostic Devices for*Bacillus* spp. Detection; Class II Special Controls Guideline for Industry and Food and Drug Administration Staff.” For availability of the guideline document, see § 866.1(e).(c) *Restriction on Distribution.* The distribution of these devices is limited to laboratories that follow public health guidelines that address appropriate biosafety conditions, interpretation of test results, and coordination of findings with public health authorities.(d) *Restriction on Use.* The use of this device is restricted to prescription use and must comply with the following:(1) The device must be in the possession of: (i)(A) A person, or his agents or employees, regularly and lawfully engaged in the manufacture, transportation, storage, or wholesale or retail distribution of such device; or (B) A practitioner, such as a physician, licensed by law to use or order the use of such device; and (ii) The device must be sold only to or on the prescription or other order of such practitioner for use in the course of his professional practice. (2) The label of the device shall bear the statement “Caution: Federal law restricts this device to sale by or on the order of a ____”, the blank to be filled with the word “physician” or with the descriptive designation of any other practitioner licensed by the law of the State in which he practices to use or order the use of the device. (3) Any labeling, as defined in section 201(m) of the Federal Food, Drug, and Cosmetic Act, whether or not it is on or within a package from which the device is to be dispensed, distributed by, or on behalf of the manufacturer, packer, or distributor of the device, that furnishes or purports to furnish information for use of the device contains adequate information for such use, including indications, effects, routes, methods, and frequency and duration of administration and any relevant hazards, contraindications, side effects, and precautions, under which practitioners licensed by law to employ the device can use the device safely and for the purposes for which it is intended, including all purposes for which it is advertised or represented. This information will not be required on so-called reminder-piece labeling which calls attention to the name of the device but does not include indications or other use information. (4) All labeling, except labels and cartons, bearing information for use of the device also bears the date of the issuance or the date of the latest revision of such labeling.

Predicate Devices

• Unmodified JBAIDS Anthrax Detection System (K051713)

Submission Summary (Full Text)

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SPECIAL 510(k): Device Modification
ODE Review Memorandum (Decision Making Document is Attached)

To: THE FILE
RE: DOCUMENT NUMBER K_071188

This 510(k) submission contains information/data on modifications made to the SUBMITTER'S own Class II, Class III or Class I devices requiring 510(k). The following items are present and acceptable (delete/add items as necessary):

1. The name and 510(k) number of the SUBMITTER'S previously cleared device.
JBAIDS Anthrax Detection Kit K051713

2. Submitter's statement that the INDICATION/INTENDED USE of the modified device as described in its labeling HAS NOT CHANGED along with the proposed labeling which includes instructions for use, package labeling, and, if available, advertisements or promotional materials (labeling changes are permitted as long as they do not affect the intended use).

3. A description of the device MODIFICATION(S), including clearly labeled diagrams, engineering drawings, photographs, user's and/or service manuals in sufficient detail to demonstrate that the FUNDAMENTAL SCIENTIFIC TECHNOLOGY of the modified device has not changed.

This change was for
The addition of the IT 1-2-3™ QFLOW™ Sample Purification kit to the JBAIDS Anthrax Detection kit.

4. Comparison Information (similarities and differences) to applicant's legally marketed predicate device including, labeling, intended use, physical characteristics, and

| | Similarities | Differences |
| --- | --- | --- |
| | Predicate | Modified |
| Intended Use | To detect target DNA sequences of B. anthracis | Same |
| Labeling | Basic package insert with no information on the IT 1-2-3QFLOW™ Sample Purification kit, ASAY-ASY | Addition under Materials Required Not Provided: IT 1-2-3QFLOW™ Sample Purification kit, ASAY-ASY-0503
IT1-2-3RNA Module, ASAY-ASY-0501
Other updates to the following sections Specimen Collection and Preparation, Sample Purification Procedure, Interfering substances, and Quality Control Purification |
| System | Detection kit | Same |
| | FLOW (Predicate) | QFLOW^{dna} (Modified) |
| --- | --- | --- |
| Lysis | 3 ml of citrated whole blood | 0.8 ml of citrated whole blood |
| Protease treatment | 400 μL of protease, 5 min @ rm temp | 60 μL of protease K, 10 μL of carrier RNA, and 600 μL of Qiagen Buffer Al @ 55-60°C for 1 hr |
| Bind DNA to filter | 9 mL of Buffer 1, centrifuge for 3 min at 3000g | Add 600 μL of ethanol and transfer 600 μL to a QIAamp spin column. Centrifuge for 1.5 min at 6000g. Repeat two more times. |
| Wash Filter | Add 9 mL of Buffer 2 and centrifuge for 3 min at 3000 g | Add 500 μL Qiagen AW1 buffer and centrifuge for 1.5 min at 6000g. Repeat with Qiagen AW2 |
| Dry Spin and Heat | Centrifuge for 5 min at 3000 g, heat the spin filter for 5 min at 88-95°C in a heat block or 70-75°C in a water bath | Centrifuge QIAamp spin column for 3 min at 12,000 g. |

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| Elute Sample | Add 1 mL of Buffer 3. Incubate for 5 min at rm temp; centrifuge for 5 min at 3000 g | Add 230 μL Qiagen Buffer AE heated to 68-72 °C. Incubate for 5 min at 68-72°C; centrifuge for 1.5 min at 6000 g. |
| --- | --- | --- |
| Dilution | Dilute purified sample 1:5 with Buffer 3 just before testing with the JBAIDS Anthrax Detection Kit | The purified sample is ready for testing with the JBAIDS Anthrax Detection Kit |

5. A Design Control Activities Summary which includes:

a) Identification of Risk Analysis method(s) used to assess the impact of the modification on the device and its components, and the results of the analysis
b) Based on the Risk Analysis, an identification of the verification and/or validation activities required, including methods or tests used and acceptance criteria to be applied
c) A declaration of conformity with design controls. The declaration of conformity should include:

i) A statement signed by the individual responsible, that, as required by the risk analysis, all verification and validation activities were performed by the designated individual(s) and the results demonstrated that the predetermined acceptance criteria were met, and
ii) A statement signed by the individual responsible, that the manufacturing facility is in conformance with design control procedure requirements as specified in 21 CFR 820.30 and the records are available for review.

The design control activities performed

a. Assay Sensitivity

Target 1, FLOW Cp 34.87 vs. QFLOW$^{\text{dna}}$ Cp 31.46

Target 2, FLOW Cp 34.68 vs. QFLOW$^{\text{dna}}$ Cp 31.76

The Cp values obtained for the QFLOW$^{\text{dna}}$ processed sample set were earlier than that obtained for the FLOW processed sample set.

b. Quantity of DNA (Spectrophotometric method)

FLOW 6.18 ng/ μL vs. QFLOW$^{\text{dna}}$ 54.61 ng/ μL

The quantity of total DNA is higher for sample processed using the QFLOW$^{\text{dna}}$ protocol.

c. OD260/OD280 ratio

FLOW 1.02 vs. QFLOW$^{\text{dna}}$ 1.96

The purity of DNA with regard to protein content is better for the QFLOW$^{\text{dna}}$ processed samples.

d. Gel Analysis of Purified DNA

The bands of the DNA purified using the QFLOW$^{\text{dna}}$ procedure were brighter, indicating the quantity of DNA is higher.

e. Removal of Inhibitory Substances

Citrated whole blood samples from 16 normal healthy donors were processed using the FLOW and the QFLOW$^{\text{dna}}$ procedures and then tested using the Target 1 and

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Target 2 assays.
Target 1, FLOW IC Cp 31.69 vs. QFLOW$^{\text{dna}}$ IC Cp 31.80
Target 2, FLOW IC Cp 31.91 vs. QFLOW$^{\text{dna}}$ IC Cp 32.04
There was adequate removal of PCR inhibitors from whole blood samples processed using the QFLOW$^{\text{dna}}$ protocol.

f. Carry-Over
Strongly positive samples (spiked at 5x $10^{6}$ CFU/mL) were purified, using FLOW and the QFLOW$^{\text{dna}}$ procedures, next to negative (unspiked) samples. Both sets of negative samples were tested by the Target 1 and Target 2 assays.
Target 1, FLOW carry-over rate 0% (0/42) vs. QFLOW$^{\text{dna}}$ 2.4% (1/42)
Target 2, FLOW carry-over rate 0% (0/42) vs. QFLOW$^{\text{dna}}$ 0% (0/42)

6. A Truthful and Accurate Statement, a 510(k) Summary or Statement and the Indications for Use Enclosure (and Class III Summary for Class III devices).

The labeling for this modified subject device has been reviewed to verify that the indication/intended use for the device is unaffected by the modification. In addition, the submitter's description of the particular modification(s) and the comparative information between the modified and unmodified devices demonstrate that the fundamental scientific technology has not changed. The submitter has provided the design control information as specified in The New 510(k) Paradigm and on this basis, I recommend the device be determined substantially equivalent to the previously cleared (or their preamendment) device.

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REVISED:3/14/95

THE 510(K) DOCUMENTATION FORMS ARE AVAILABLE ON THE LAN UNDER 510(K) BOILERPLATES TITLED "DOCUMENTATION" AND MUST BE FILLED OUT WITH EVERY FINAL DECISION (SE, NSE, NOT A DEVICE, ETC.).

"SUBSTANTIAL EQUIVALENCE" (SE) DECISION MAKING DOCUMENTATION

K071188

Reviewer: Alexandra Wong/Freddie Poole

Division/Branch: DMD/BACB

Device Name: JBAIDS Anthrax Detection kit

Product To Which Compared (510(K) Number If Known): k051713

YES NO

| 1. Is Product A Device | x | | If NO = Stop |
| --- | --- | --- | --- |
| 2. Is Device Subject To 510(k)? | x | | If NO = Stop |
| 3. Same Indication Statement? | x | | If YES = Go To 5 |
| 4. Do Differences Alter The Effect Or Raise New Issues of Safety Or Effectiveness? | | | If YES = Stop NE |
| 5. Same Technological Characteristics? | x | | If YES = Go To 7 |
| 6. Could The New Characteristics Affect Safety Or Effectiveness? | | | If YES = Go To 8 |
| 7. Descriptive Characteristics Precise Enough? | x | | If NO = Go To 10
If YES = Stop SE |
| 8. New Types Of Safety Or Effectiveness Questions? | | | If YES = Stop NE |
| 9. Accepted Scientific Methods Exist? | | | If NO = Stop NE |
| 10. Performance Data Available? | | | If NO = Request Data |
| 11. Data Demonstrate Equivalence? | | | Final Decision: SE |

Note: In addition to completing the form on the LAN, "yes" responses to questions 4, 6, 8, and 11, and every "no" response requires an explanation.

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1. Intended Use:

The JBAIDS Anthrax Detection System is a real-time polymerase chain reaction (PCR) test system intended for the qualitative *in vitro* diagnostic (IVD) detection of target DNA sequences on the pXO1 plasmid (Target 1) and the pXO2 plasmid (Target 2) from *Bacillus anthracis*. The system can be used to test human whole blood collected in sodium citrate from individuals suspected of having anthrax, positive blood cultures, and cultured organisms grown on blood agar plates. The JBAIDS Anthrax Target 2 assay is used as a supplementary test only after a positive result with the Target 1 Assay.

JBAIDS Anthrax Target 1 and Target 2 Assays are run on the JBAIDS instrument using the Diagnostic Wizard.

2. Device Description: Provide a statement of how the device is either similar to and/or different from other marketed devices, plus data (if necessary) to support the statement. Does the device design use software? Is the device sterile? Is the device for single use? Is the device over-the-counter or prescription use? Does the device contain drug or biological product as a component? Is this device a kit? Provide a summary about the devices design, materials, physical properties and toxicology profile if important.

The Joint Biological Agent Identification and Diagnostic System (JBAIDS) Anthrax Detection System is a fully integrated *in-vitro* diagnostic (IVD) system composed of the JBAIDS instrument with laptop computer, software, 2 different freeze-dried reagent assays (in one kit) for the qualitative detection of pathogenic *Bacillus anthracis*, and 4 different sample preparation protocols, two for isolating target DNA from whole blood, one for processing blood culture, and another for processing colonies. This submission is for a modification to the sample purification kit described below.

Device Modification

The modification consists in the addition of the QFLOW$^{\text{dna}}$ Sample Purification Kit for the purification of whole blood samples. The QFLOW$^{\text{dna}}$ protocol offers the advantages of requiring a smaller sample volume, and all processing steps can be performed using a smaller bench top centrifuge. The table below exhibits a summary of changes to the modified kit.

| | FLOW (Predicate) | QFLOW$^{\text{dna}}$ (Modified) |
| --- | --- | --- |
| Lysis | 3 ml of citrated whole blood | 0.8 ml of citrated whole blood |
| Protease treatment | 400 μL of protease, 5 min @ rm temp | 60 μL of protease K, 10 μL of carrier RNA, and 600 μL of Qiagen Buffer Al @ 55-60°C for 1 hr |
| Bind DNA to filter | 9 mL of Buffer 1, centrifuge for 3 min at 3000g | Add 600 μL of ethanol and transfer 600 μL to a QIAamp spin column. Centrifuge for 1.5 min at 6000g. Repeat two more times. |
| Wash Filter | Add 9 mL of Buffer 2 and centrifuge for 3 min at 3000 g | Add 500 μL Qiagen AW1 buffer and centrifuge for 1.5 min at 6000g. Repeat with Qiagen AW2 |
| Dry Spin and Heat | Centrifuge for 5 min at 3000 g, heat the spin | Centrifuge QIAamp spin column for 3 min at |

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| | filter for 5 min at 88-95°C in a heat block or 70-75°C in a water bath | 12,000 g. |
| --- | --- | --- |
| Elute Sample | Add 1 mL of Buffer 3. Incubate for 5 min at rm temp; centrifuge for 5 min at 3000 g | Add 230 μL Qiagen Buffer AE heated to 68-72 °C. Incubate for 5 min at 68-72°C; centrifuge for 1.5 min at 6000 g. |
| Dilution | Dilute purified sample 1:5 with Buffer 3 just before testing with the JBAIDS Anthrax Detection Kit | The purified sample is ready for testing with the JBAIDS Anthrax Detection Kit |

## EXPLANATIONS TO "YES" AND "NO" ANSWERS TO QUESTIONS ON PAGE 1 AS NEEDED

11. Explain how the performance data demonstrates that the device is or is not substantially equivalent:

The sponsor included all required information in the submission to meet the criteria for a Special 510(k). The sponsor submitted descriptions with explanations of the device modification, i.e., the addition of a new sample purification kit; and a summary of their design control validation studies.

ATTACH ADDITIONAL SUPPORTING INFORMATION