Contains Nonbinding Recommendations 
 
 
Content and Format for Abbreviated 
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510(k)s for Early Growth Response 1 
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(EGR1) Gene Fluorescence In-Situ 
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Hybridization (FISH) Test System for 
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Specimen Characterization Devices 
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6
Guidance for Industry and Food and 
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Drug Administration Staff  
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9
Document issued on June 17, 2015.  
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The draft of this guidance was issued on September 26, 2014. 
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14 For questions about this document, contact the Division of Molecular Genetics and Pathology 
15 at 301-796-6179 or Shyam Kalavar at 301-796-6807 or at Shyam.Kalavar@fda.hhs.gov.  
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20 U.S. Department of Health and Human Services 
21  Food and Drug Administration 
22  Center for Devices and Radiological Health 
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24  Office of In Vitro Diagnostics and Radiological Health 
25  Division of Molecular Genetics and Pathology 
26  Molecular Genetics Branch 
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30 Contains Nonbinding Recommendations 
 
 
Preface 
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Public Comment 
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35 You may submit electronic comments and suggestions at any time for Agency consideration to 
36 http://www.regulations.gov . Submit written comments to the Division of Dockets 
37 Management, Food and Drug Administration, 5630 Fishers Lane, Room 1061, (HFA-305), 
38 Rockville, MD 20852. Identify all comments with the docket number FDA-2014-D-1242. 
39 Comments may not be acted upon by the Agency until the document is next revised or 
40 updated. 
41
Additional Copies 
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43  
44 Additional copies are available from the Internet.  You may also send an e-mail request to 
45 CDRH-Guidance@fda.hhs.gov to receive a copy of the guidance. Please use the document 
46 number 1400030 to identify the guidance you are requesting. 
47
48 Contains Nonbinding Recommendations 
 
 
Table of Contents 
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51 I.  Introduction ...................................................................................................................... 1 
52 II.  Scope ................................................................................................................................ 1 
53 III.  Policy ............................................................................................................................... 2 
54 A.  Content and Format of an Abbreviated 510(k) .......................................................... 2 
55 1.  Coversheet .......................................................................................................... 2 
56 2.  Proposed labeling ................................................................................................ 2 
57 3.  Summary report .................................................................................................. 2 
58 4.  Device Information ............................................................................................. 3 
59 5.  Device Performance Specifications .................................................................... 4 
60 6.  Clinical Validity .................................................................................................. 6 
61 B.  Labeling ...................................................................................................................... 7 
62 Contains Nonbinding Recommendations 
Content and Format for Abbreviated 
63
510(k)s for Early Growth Response 1 
64
(EGR1) Gene Fluorescence In-Situ 
65
Hybridization (FISH) Test System for 
66
Specimen Characterization Devices  
67
68
Guidance for Industry and Food and Drug 
69
Administration Staff  
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71  
72 This guidance represents the current thinking of the Food and Drug Administration (FDA 
73 or Agency) on this topic.  It does not establish any rights for any person and is not binding 
74 on FDA or the public.  You can use an alternative approach if it satisfies the requirements 
75 of the applicable statutes and regulations.  To discuss an alternative approach, contact the 
76 FDA staff responsible for this guidance as listed on the title page. 
77  
I. Introduction 
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80 FDA is issuing this guidance to provide industry and agency staff with recommendations for 
81 the suggested format and content of an Abbreviated 510(k) submission for early growth 
82 response 1 (EGR1) gene fluorescence in-situ hybridization (FISH) test system for specimen 
83 characterization devices and recommendations for addressing certain labeling issues relevant 
84 to the review process specific to these devices.   
85  
86 FDA's guidance documents, including this guidance, do not establish legally enforceable 
87 responsibilities.  Instead, guidances describe the Agency's current thinking on a topic and 
88 should be viewed only as recommendations, unless specific regulatory or statutory 
89 requirements are cited.  The use of the word should in Agency guidances means that 
90 something is suggested or recommended, but not required.  
91  
II.  Scope
92  
93 The scope of this document is limited to the devices identified in 21 CFR 864.1870 as: 
94 An early growth response 1 (EGR1) gene fluorescence in-situ hybridization (FISH) test 
95 system for specimen characterization is a device intended to detect the EGR1 probe target on 
96 chromosome 5q in bone marrow specimens from patients with acute myeloid leukemia 
97 (AML) or myelodysplastic syndrome (MDS).  The assay results are intended to be 
98 interpreted only by a qualified pathologist or cytogeneticist.  These devices do not include 
  1  Contains Nonbinding Recommendations 
99 automated systems that directly report results without review and interpretation by a 
100 qualified pathologist or cytogeneticist.  These devices also do not include any device 
101 intended for use to select patient therapy, predict patient response to therapy or to screen for 
102 disease as well as any device with a claim for a particular diagnosis, prognosis, monitoring or 
103 risk assessment. 
104
III.  Policy  
105
106
107 The following are the recommended content and format of an abbreviated 510(k) for these 
108 devices and recommendations for addressing certain labeling issues relevant to the review 
109 process specific to these devices. 
110
A. Content and Format of an Abbreviated 510(k) 
111
112
113 An Abbreviated 510(k) submission must include the required elements identified in 21 CFR 
114 807.87, including the proposed labeling for the device sufficient to describe the device, its 
115 intended use, and the directions for its use.  In an Abbreviated 510(k) for this device, FDA 
116 may consider the contents of a summary report to be appropriate supporting data within the 
117 meaning of 21 CFR 807.87(f) or (g); therefore, we recommend that you include a summary 
118 report.  The summary report should describe how this guidance document was used during 
119 the device development and testing and the methods or tests used.  The report should also 
120 include a summary of the test data or description of the acceptance criteria applied to address 
121 the risks identified in this document, as well as any additional risks specific to your device.  
122 This section provides suggestions about how to compose your Abbreviated 510(k) 
123 submission, including a suggested order and headings. 
124  
125 1.  Coversheet 
126 The coversheet should prominently identify the submission as an Abbreviated 510(k) 
127 and cite the title of this guidance document. 
128
129 2.  Proposed labeling 
130 Include proposed labels, labeling, and advertisements sufficient to describe the 
131 device, its intended use, and the directions for its use.  21 CFR 807.87(e).  Refer to 
132 the section titled Labeling for specific information that you should include in the 
133 labeling for this type of device. 
134  
135 3.  Summary report 
136 The special controls for an Early Growth Response 1 (EGR1) Gene Fluorescence in-
137 situ Hybridization (FISH) Test System for Specimen Characterization have been 
138 outlined in regulation 21 CFR 864.1870.  The Abbreviated 510(k) submission should 
139 include a summary report in tabular format that contains the information required by 
140 these special controls as well as confirmation that the studies have been conducted 
141 and met the appropriate pre-specified acceptance criteria.  These studies are listed 
142 below with additional explanatory information intended to provide clarity about the 
143 content of the submission. 
  2  Contains Nonbinding Recommendations 
144
145 4.  Device Information 
146
147 a.  Device Intended Use/Indications for Use statement:  You should provide a Device 
148 Intended Use/Indications for Use statement. 
149  
150 b.  Description of all probes included in the kit:  You must provide a detailed 
151 description of all probes included in the kit.  21 CFR 864.1870(b)(1)(i).  The 
152 information you provide should include, but not necessarily be limited to, the 
153 identification of labeling moiety, the chromosome specificity (e.g., chromosome, 
154 band), and the nature of probes (e.g., break-apart, dual color, dual fusion).  
155   
156 c.  Purpose of Each Probe:  You must provide the purpose of each probe.  21 CFR 
157 864.1870(b)(1)(ii).  You should indicate the chromosomal abnormality the probe 
158 was designed to detect (e.g., deletion/amplification).  You should use standard 
159 scientific nomenclature and provide a glossary of terms where relevant. 
160  
161 d.  Probe Molecular Specificity:  You must provide probe molecular specificity.  21 
162 CFR 864.1870(b)(1)(iii).  You should provide end-sequencing information for each 
163 probe and its link to the reference human genome sequence. 
164  
165 e.  Probe Specificity:  You must provide probe specificity.  21 CFR 
166 864.1870(b)(1)(iv). You should provide evidence of specific binding to the 
167 expected chromosomal site (chromosome and band) for 5 or more metaphase 
168 samples from different normal individuals (indicate sex of each sample) and for 20 
169 or more consecutive intact metaphases for each sample.  When more than one 
170 site/technologist is used, each should perform a complete analysis of the 5 samples 
171 and 20 or more consecutive intact metaphase stated above.  You should list all 
172 results and annotate number and location of unexpected signals.  Provide specificity 
173 calculations with 95% confidence intervals.  
174
175   f.  Probe Limits:  You must provide probe limits.  21 CFR 864.1870(b)(1)(v).  You 
176 should indicate the highest and lowest probe concentrations that produce acceptable 
177 signals.   
178
179   g.  Probe Sensitivity:  You must provide probe sensitivity.  21 CFR 864.1870(b)(1)(vi).  
180 You should provide expected and observed signal count for 25 samples from 
181 different normal individuals (indicate sex), counting 200 consecutive scoreable 
182 nuclei from each sample.  You should provide sensitivity calculations with 95% 
183 confidence intervals.  You should describe expected values (cut-off) and provide 
184 calculations. 
185
186   h.  Reagents:  You must provide a specification of the devices required ancillary 
187 reagents, instrumentation and equipment.  21 CFR 864.1870(b)(1)(vii). 
188
  3  Contains Nonbinding Recommendations 
189   i.  Pre-Analytics: You must provide a specification of the specimen collection, 
190 processing, storage and slide preparation methods.  21 CFR 864.1870(b)(1)(viii).  
191
192   j.  Assay Procedure:  You must provide a specification for the assay procedure.  21 
193 CFR 864.1870(b)(1)(ix).  You should provide a detailed description of the 
194 principles of operation, including the procedure for detecting and differentiating 
195 multiple analytes (if applicable).   
196
197 k.  Controls and Risk Mitigation:  You must provide a specification for the control 
198 elements that are incorporated into the recommended testing procedures and 
199 specification of risk mitigation elements (description of all additional procedures, 
200 methods, and practices incorporated into the directions for use that mitigate risks 
201 associated with testing).  21 CFR 864.1870(b)(1)(x and xi).  You should describe the 
202 testing conditions, procedures that use the controls designed to safeguard against 
203 conditions that can cause false positive and false negative results.  These should 
204 include at a minimum: 
205
206 i.  Description of, or recommendations for, any internal controls.  
207 ii.  Features and additional controls that monitor procedural errors or factors 
208 (e.g., degradation of the master mix) that adversely affect performance of 
209 the test. 
210
211 l.  Criteria for Interpreting Test Results and Reporting: You must provide a 
212 specification of the criteria for test result interpretation and reporting.  21 CFR 
213 864.1870(b)(1)(xii).  You should describe how positive, negative, equivocal (if 
214 applicable), or invalid results are determined and how they should be interpreted 
215 including slide adequacy, signal enumeration, expected values and results 
216 interpretation. 
217
218 5.  Device Performance Specifications 
219 The performance studies supporting the submission must at a minimum include: 1) 
220 Device analytical sensitivity data, 2) Device analytical specificity data, 3) Device 
221 reference limit data, 4) Device precision/reproducibility data, and 5) Device stability data 
222 to include:  
223 A) Real-time Stability  
224 B) Freeze-Thaw Stability  
225 C) Transport and Temperature Stability  
226 D) Post-Hybridization Signal Stability  
227 E) Photostability of Probe  
228
229 21 CFR 864.1870(b)(1)(xiii-xvii)).  We recommend providing a summary of this 
230 information in a tabular format (See Table 1 for an example of how the information 
231 may be formatted) that includes the protocol, pre-specified acceptance criteria, 
232 justification for the pre-specified acceptance criteria, and whether the pre-specified 
233 acceptance criteria were met for the performance studies. 
234
  4  Contains Nonbinding Recommendations 
235 Table 1. 
Performance  Protocol & Pre- Justification for Pre- Testing Confirming Pre-
Study  Specified Acceptance  Specified Acceptance  Specified Acceptance Criteria 
Criteria  Criteria  Met 
Precision:  Describe the specimen  Provide justification for the  Indicate whether the pre-
Intra-Day and   panel   pre-specified acceptance  specified acceptance criteria 
Inter-Day   Note: This study is  criteria  were met and whether any 
expected to include  results were excluded. No 
minimally 2 high  results should be excluded 
positive specimens, 2  without satisfactory 
low positive specimens  justification.  
and 2 negative 
specimens. 
 
Describe the study 
protocol  
  
Indicate how many lots 
were included in the 
study  
 
Indicate how the results 
were analyzed, e.g, red 
and green signal patterns 
of 200 nuclei evaluated 
by 2 technologists, 
where each technologist 
evaluated 100 nuclei per 
panel member 
 
Indicate the pre-
specified acceptance 
criteria 
Reproducibility: 
Same as above 
Inter-Site  
Lot to Lot 
Same as above 
Reproducibility 
Real-Time 
Same as above, and also 
Stability 
indicate the attributes 
evaluated. 
In-Use Freeze-
Same as above, and also 
Thaw Stability 
indicate the attributes 
evaluated. 
Transport and 
Same as above, and also 
Temperature 
indicate the attributes 
Extreme 
evaluated. 
Stability 
Post-
Same as above, and also 
hybridization 
indicate the attributes 
Signal Stability 
evaluated. 
  5  Contains Nonbinding Recommendations 
Probe Photo-
Same as above, and also 
stability 
indicate the attributes 
evaluated. 
236
237 Patient samples derived from the intended use population (e.g., patients with acute 
238 myeloid leukemia or myelodysplastic syndrome) should be used in these studies.  
239 When this is not possible, spiked normal samples or samples derived from 
240 representative positive and negative cultured cells may be appropriate; however, 
241 using spiked or cultured cell samples as the only matrix in the evaluation may not 
242 provide an accurate assessment of the performance characteristics.  Appropriate assay 
243 controls should be used when conducting the performance studies.  
244
245 6.  Clinical Validity 
246 You must include documentation that demonstrates the clinical validity of the device.  
247 21 CFR 864.1870(b)(1)(xxviii).  The documentation must include data from clinical 
248 studies, a minimum of two peer-reviewed published literature references using the 
249 specific device seeking marketing clearance, or both. 21 CFR 864.1870(b)(1)(xiii-
250 xvii).  Documentation for the clinical studies and peerreviewed published literature 
251 references cited must include the following elements:  
252     A) Documentation that the sponsors probe was used in the literature reference  
253     B) Number & type of specimens  
254     C) Target population studied  
255     D) Upper reference limit  
256     E) Range of positive probe results  
257
258 21 CFR 864.1870(b)(1)(xiii-xvii).  The information should be summarized in tabular 
259 format (See Table 2 for an example of how the information may be formatted).  
260  
261 If you use peer-reviewed published scientific literature references to support the 
262 510(k) submission, then a declaration should be provided in the Abbreviated 510(k) 
263 submission that the literature supports the devices claims.  Supportive peer-reviewed 
264 published literature should use the same product for which you are seeking clearance 
265 and contain valid safety and effectiveness data.  Any unpublished data safety and 
266 effectiveness data that you have for your device should be provided in the 510(k) 
267 submission.  You should cite only relevant published literature for the defined clinical 
268 setting where product was used and identify specimen matrices.   
269  
270 Table 2. 
Conditions  Data Source 1  Data Source 2  Data Source 3 
Authors name, et al.  Authors name, et.al  Authors name, et al. 
Was the specific device 
under review in the 
Yes  Yes  Yes 
submission used in the 
study? 
Was the specimen type 
in the study  Yes  Yes  Yes 
representative of the 
  6  Contains Nonbinding Recommendations 
claimed specimen 
type(s) 
Target population  Indicate the diseased 
(disease status)  population(s) in the 
paper, e.g., known or 
suspected del(5q) 
having MDS or AML 
Upper reference limit  Indicate the percentage 
(percentage and per  of relevant nuclei used 
200 nuclei)  as a clinical decision 
point, e.g., , 6% or 12 
1R2G patterns per 200 
scoreable interphase 
nuclei 
Total Number of  Indicate the number of 
specimens tested for  bone marrow and or 
each claimed type   peripheral blood 
specimens 
Number of specimens 
with a positive probe  Indicate the number  N 
N 
result [5q- (1R2G)]  (N) per disease state 
Range of positive probe 
results 
271
B. Labeling 
272
273
274 The 21 CFR 809.10(b)(12) compliant labeling must include a statement summarizing the 
275 data identified in 21 CFR 864.1870(b)(1)(xiii)-(xviii) and a description of the studies 
276 supporting the information, including the pre-specified acceptance criteria for these 
277 performance studies, justification for the pre-specified acceptance criteria, and whether the 
278 pre-specified acceptance criteria were met.  21 CFR 864.1870(b)(2). 
279
280 The 21 CFR 809.10 compliant labeling must include:  
281
282 i)  A warning that reads The assay results are intended to be interpreted only by a 
283 qualified pathologist or cytogeneticist.  
284 ii)  A warning that reads This device is not for high-risk uses such as selecting therapy, 
285 predicting therapeutic response or disease screening. 
286 iii)  A warning that reads The use of this device for diagnosis, monitoring or risk 
287 assessment has not been established.  
288
289 21 CFR 864.1870(b)(3).  The labeling should also include specific instructions and the 
290 clinical training needed for the safe use of the device.   
291
292 If peer-reviewed published scientific literature references are used to support the Abbreviated 
293 510(k) submission, then you should include a statement in the labeling that reads "Cited 
294 literature may discuss device uses that have not been approved or cleared by FDA."    
  7