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1 From the Department of Radiology, University of Michigan Hospital, UH B1F510, 1500 E Medical Center Dr, Ann Arbor, MI 48109-0030. From the 1997 RSNA scientific assembly. Received August 10, 1998; revision requested September 8; revision received November 30; accepted January 21, 1999. Supported in part by United States Public Health Service grant CA 48129 and by U.S. Army Medical Research and Materiel Command grant DAMD 17-96-1-6254. B.S. supported by Career Development award DAMD 17-96-1-6012 from the U.S. Army Medical Research and Materiel Command. N.P. supported by a grant from the Whitaker Foundation. Address reprint requests to H.P.C. (e-mail: chanhp@umich.edu).
| Abstract |
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MATERIALS AND METHODS: The authors previously developed an automated computer program for estimation of the relative malignancy rating of masses. In the present study, the authors conducted observer performance experiments with receiver operating characteristic (ROC) methodology to evaluate the effects of computer estimates on radiologists' confidence ratings. Six radiologists assessed biopsy-proved masses with and without CAD. Two experiments, one with a single view and the other with two views, were conducted. The classification accuracy was quantified by using the area under the ROC curve, Az.
RESULTS: For the reading of 238 images, the Az value for the computer classifier was 0.92. The radiologists' Az values ranged from 0.79 to 0.92 without CAD and improved to 0.870.96 with CAD. For the reading of a subset of 76 paired views, the radiologists' Az values ranged from 0.88 to 0.95 without CAD and improved to 0.930.97 with CAD. Improvements in the reading of the two sets of images were statistically significant (P = .022 and .007, respectively). An improved positive predictive value as a function of the false-negative fraction was predicted from the improved ROC curves.
CONCLUSION: CAD may be useful for assisting radiologists in classification of masses and thereby potentially help reduce unnecessary biopsies.
Index terms: Breast neoplasms, 00.31, 00.32 Breast neoplasms, radiography, 00.111, 00.119 Breast radiography, 00.111, 00.119 Computers, diagnostic aid Receiver operating characteristic curve (ROC)
| Introduction |
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Computer-aided diagnosis (CAD) is considered to be one of the approaches that may improve the efficacy of mammography (4). With CAD, a computerized detection algorithm alerts a radiologist to the location of the suspicious lesions, and/or a trained computer classifier provides the radiologist with an estimate of the likelihood of malignancy of a lesion. The radiologist takes into consideration the information provided by the computer before making a decision. This "second opinion" may improve the diagnostic accuracy because it serves as a form of double reading (5). Furthermore, a computer evaluation is often more consistent and reproducible than a human decision maker (6).
Considerable research has been devoted to the development of computerized schemes for the detection and classification of mammographic abnormalities. These efforts have advanced the CAD technology such that clinical application appears to be possible in the near future. It is, therefore, necessary to evaluate the effects of CAD on radiologists' detection and diagnosis of mammographic lesions. In a previous receiver operating characteristic (ROC) study, we demonstrated that CAD could improve radiologists' accuracy in the detection of subtle microcalcifications on mammograms (7). Kegelmeyer et al (8) also reported an improvement in radiologists' sensitivity for the detection of spiculated masses with use of a computer aid. For the classification of mammographic lesions, it has been shown that a computer classifier that estimated the likelihood of malignancy on the basis of mammographic features extracted by radiologists could improve radiologists' accuracy in distinguishing malignant from benign lesions (911).
We previously conducted ROC studies to compare the performance of radiologists with that of the computer (12) and to compare radiologists' ability to classify masses with and without CAD (13). Jiang et al (14) also performed an ROC study of the effect of CAD on radiologists' performance in classifying microcalcifications. The results of all of these observer performance studies indicate the potential to improve mammographic interpretation with a computer aid.
We have developed an automated method to analyze masses seen on mammograms (1517). A mass is segmented from its surrounding breast tissue, and an image transformation technique is used to transform the mass margin from the polar coordinate system to the Cartesian coordinate system. A linear discriminant classifier then extracts the useful texture features from the transformed image and merges them into a relative malignancy rating. Our approach is different from others that use a trained classifier to merge radiologist-extracted image features or feature codes by using the American College of Radiology Breast Imaging Reporting and Database System lexicon (911). Our fully automated method has the advantage that, unlike a human reader, it does not have variability in feature recognition and coding. In addition, the computer may be able to extract some information, such as texture features, that may not be readily perceived by human eyes. We conducted an ROC study to evaluate whether this computer aid can improve radiologists' performance in the classification of mammographic masses (13). The results of our observer performance study are described in this article.
Other investigators also have reported on automated algorithms for the classification of mammographic masses (1821). The methods used in these algorithms varied, and their accuracy in classification cannot be compared directly because of the differences in the data sets. However, the effects of CAD on radiologists' performance are not expected to depend strongly on the specific algorithm if different computer aids of comparable accuracy are used. Therefore, the applications of the findings of this study should not be limited to our computerized classification aid.
| MATERIALS AND METHODS |
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The mammograms were acquired with a contact technique. The dedicated mammographic systems had a molybdenum anode and molybdenum filter, a 0.3-mm nominal focal spot, and a reciprocating grid. MinR/MinR-E screen-film systems (Eastman-Kodak, Rochester, NY) were used with these units. Sixty-two of the malignant masses and six of the benign masses were judged to be spiculated by a radiologist (M.A.H.) experienced in mammography. The radiologist also measured the size (ie, longest dimension) and ranked the visibility of the masses on a scale of 1 (obvious) to 5 (subtle) relative to the range of visibility of masses encountered in clinical practice. For a description of the masses included in the data set, histograms of the size and visibility of the masses are shown in Figures 1a and 1b, respectively.
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For the observer experiments, we used laser-printed images of the digitized mammograms for all readings. The images were printed with a 969HQ laser imager (Imation, Oakdale, Minn) that was connected to a Macintosh computer (Apple Computer, Cupertino, Calif) through a special digital interface. The interface provided a 12-bit in, 10-bit out look-up table and allowed images to be scaled to different factors with 15 interpolation methods. Because this laser imager has a pixel size of about 0.085 mm, we enlarged the images by about 18% during printing to maintain them at the same size as the original mammograms. One of the interpolation methods was chosen by an experienced radiologist (M.A.H.), who inspected the printed images with a magnifier and evaluated the sharpness of the spicules and mass boundaries. Because of the small pixel size used for both digitization and printing, basically no noticeable blurring of the masses could be seen with the chosen interpolation method. The images were also inspected for the potential contouring effect of 10-bit output images, but no noticeable artifacts could be found. A linear pixel valuetooutput optical density calibration curve of the laser imager was used for the printing. All images were printed with the same settings.
Computerized Classification of Masses
Our computerized method of classifying mammographic masses has been described in detail previously (1517). The method is summarized as follows: A region of interest that contained the biopsy-proved mass was identified on the mammogram by the radiologist. Background correction based on a distance-weighted estimation method was applied to the region of interest to reduce the low-frequency density variation in the region. A median-filtered smoothed image and two high-frequency enhanced images were generated from the background-corrected region of interest. The smoothed and enhanced gray-level values at each pixel were used as features in a k-means clustering algorithm to classify the pixels into two clusters; one was the mass, and the other was the surrounding breast tissue background. By choosing an appropriate criterion, a mass region slightly smaller than the actual mass that was visible on the image was segmented.
The boundary of the segmented region was smoothed by morphologic filtering. A new image transformation technique, referred to as the rubber-band-straightening transform, was used to transform a 40-pixel-wide region that surrounded the segmented mass boundary into a rectangular region. After transformation, the mass margin became approximately parallel, and any spicules that were radiating from the mass became approximately perpendicular, to the long dimension of the rectangular region. The rubber-band-straightening transform enabled the spicules to be aligned approximately in a uniform direction and thus facilitated the extraction of texture features from the margin of the mass. An example of a rubber-band-straighteningtransformed image is shown in Figure 2.
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,d(i,j) is the joint probability of the occurrence of gray levels i and j for pixel pairs that are separated by a distance d and at a direction
(22). For analysis of the masses, the spatial gray-level dependence matrices were constructed for 10 pixel distances (d = 1, 2, 3, 4, 6, 8, 10, 12, 16, 20 pixels) and in four directions (0°, 45°, 90°, 135°) relative to the mass boundary. Therefore, a total of 320 spatial gray-level dependence texture features were extracted. The second set of texture features was derived from the run length statistics matrices of the horizontal and vertical gradient images of the rubber-band-straighteningtransformed margin region. Five texture measures were extracted from the run length statistics matrix in each of the two directions (0° or 90°) on each gradient image. A total of 20 run length statistics texture features were thus obtained. Therefore, we had a total of 340 features from the two types of texture measures.
A stepwise linear discriminant feature selection procedure (23) was used to select the most effective features from the available feature set. A total of 41 features were selected. The selected features were input into the Fischer linear discriminant classifier (24) as predictor variables. A "leave one case out" resampling scheme was used to train and test the classifier. A histogram illustrating the test discriminant scores of the 253 masses is shown in Figure 3. For this classifier, a smaller discriminant score corresponded to a higher likelihood of malignancy. By using the test discriminant score as the decision variable, the performance of the computer classifier could be evaluated by using ROC analysis (17,25,26) and compared with that of the radiologists, as described later.
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The purpose of the transformation was to provide a simple and intuitive relative scale for the observer. Because the transformation was linear and monotonic, the distributions of the normal and abnormal samples, as well as their ROC curves, were not affected, with the exception of a small error caused by making the decision variables discrete. Furthermore, the slope a and intercept b parameters that were fitted to the transformed discriminant scores for the normal and abnormal samples by using the LABROC program (26) were used to generate a binormal distribution. The fitted binormal distribution with the relative malignancy rating on a 110 scale (Fig 4), together with the computer's ROC curve, were shown and explained to the observers during a training session.
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There were two reading sessions in each experimentone with CAD and the other without CAD. The observers were asked to rate the likelihood of malignancy of the masses on a 10-point confidence rating scale under all reading conditions. In the first session, half the observers interpreted the images without CAD, and the other half interpreted them with CAD. The two reading sessions in the same experiment were separated by at least 3 weeks, and the two experiments were separated by 6 months. For all four reading sessions, the observer had unlimited time to read each case. To estimate the average reading time per case for each observer, the reading time for each case was recorded by using a stopwatch.
In the first experiment, the data set of 253 single-view mammograms was divided into a training set of 15 mammograms and a study set of 238 mammograms (117 benign, 121 malignant). In each reading session, training was conducted before the reading of the study images. For the reading session with CAD, the fitted binormal distributions of the computer rating scores (Fig 4) for the entire data set were explained to the observer during training to familiarize the observer with the computer's rating scale. The computer rating of the mass was displayed on each image. After reading each training image, the observer was told the results of biopsy of the mass.
Each observer read the entire data set in one reading session. The order of the study images was randomized by a random number generator. The random sequence was different for each observer and for each reading session by the same observer. For the reading session with CAD, the observer was free to look at the computer rating, which was displayed on the image, either before or after estimating the likelihood of malignancy of the mass. However, each observer was asked to always read the computer rating before making a final decision. The observer was not informed of the pathologic results of any mass on the study images.
The second experiment was very similar to the first experiment. From the 238 single-view mammograms, 76 matched pairs (37 benign, 39 malignant) of craniocaudal and mediolateral oblique or lateral views were found. Another six pairs of two-view mammograms were identified from the rest of the images and used as training cases. The remaining mammograms were either single-view images or additional views of the pairs already chosen, so they were not used in this experiment. In this experiment, the observers were not informed of the pathologic results of any study case in any reading session. The 76 pairs of mammograms were read in one reading session by each observer.
For the reading session with CAD, the rating of the mass in each view was displayed on the respective image. The computer ratings of the mass on the two views were generally different. It was up to the observer to decide how to merge the two-view information. Observers were asked to give a single rating of the mass after reading both views.
ROC Analysis
The confidence ratings of each observer obtained from each reading condition were analyzed by using ROC methodology, and the classification accuracy was quantified by using the area under the ROC curve, Az. A maximum likelihood estimation of the binormal distribution was fitted to the confidence ratings by using the LABROC program. This program provides an estimate of the Az and of the a and b parameters of the ROC curve. The statistical significance of the difference in Az between the reading with CAD and that without CAD was estimated with two methods: One was the Student paired t test for observer-specific paired data; the other was the Dorfman-Berbaum-Metz method for analysis of multireader, multicase ROC data (28). The statistical significance of the difference in Azfor reading single-view and two-view mammograms was estimated by using the Student paired t test for the six observers. The Student paired t test takes into account the statistical variation of readers, whereas the Dorfman-Berbaum-Metz method considers both reader variation and case sample variation by means of an analysis of variance approach. Therefore, the results of Dorfman-Berbaum-Metz analysis can be generalized to the population of readers as well as to the population of case samples.
Positive Predictive Value
An ROC curve represents the entire range of operating conditions of a diagnostic process and is independent of disease prevalence. When the disease prevalence is known, any operating point on an ROC curve can be used to derive the PPV and the corresponding false-negative fraction (false-negative fraction = 1 - true-positive fraction) on the basis of the following relationship: PPV = TPF x P(M)/[TPF x P(M) + FPF x P(B)], where TPF is the true-positive fraction, FPF is the false-positive fraction at the chosen decision threshold, and P(M) and P(B) are the prevalences of malignant and benign cases, respectively. By varying the decision threshold, the dependence of the PPV on the false-negative fraction can be derived.
Because our data set did not include masses on which biopsy had not been performed, the ROC curves obtained in this study cannot be generalized to predict the performance of the computer classifier and radiologists in clinical practice. However, to demonstrate the possible effect of CAD on the PPV in the population of masses in which biopsy is likely to be performed under the current clinical criteria, we can estimate the PPV by using the prevalence of the malignant and benign masses in this patient group. Because the PPV of masses sent for biopsy ranges from about 25% to 44% in general and from about 25% to 30% at our institution, for the purposes of our estimation, we assumed that the P(M) was 25% and the P(B) was 75% in this population. A higher prevalence of malignant cases would cause an increase in the PPV, but the trend between the PPV curves with and without CAD would be similar.
| RESULTS |
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A trend similar to that with the single-view readings was observed with the two-view readings. The Az value of the computer classifier for the corresponding 152 single-view masses was 0.91 ± 0.02. The classification accuracy of all six radiologists improved when they read the mammograms with the computer aid. The increase in the Az values ranged from 0.01 to 0.07. The improvement was statistically significant (P = .007, Student paired t test; P = .026, Dorfman-Berbaum-Metz method). With CAD, two radiologists achieved an Az value of 0.97, which was higher than that obtained by the radiologists alone or by the computer alone. These results indicate that the second opinion provided by the computer classifier might have strengthened the radiologists' confidence in the interpretation of some difficult cases but had less influence on the radiologists' decision when the computer made mistakes or when the radiologists were confident about their decision.
As can be seen from the data in Table 1, the radiologists' accuracy in classifying masses by reading two-view mammograms was consistently higher than that by reading single-view mammograms (P = .008). This trend remained when they read the mammograms with CAD (P = .007). These findings are consistent with the clinical experience of the radiologists that at least two views of mammograms are needed to effectively evaluate a suspicious lesion.
The PPV as a function of the false-negative fraction was derived from the fitted ROC curves under the assumption that the prevalence of malignant masses was 25% in the population of masses sent for biopsy. The PPVs estimated for the six observers who read the two-view mammograms with and without CAD are plotted in Figure 9. CAD would provide an improvement in the PPV in the high false-negative fraction range for all observers except readers 2 and 5. The increase in the PPV at a decision threshold of "no missed malignant mass" (ie, false-negative fraction = 0) varied over a wide range; the largest gain, 39%, would be achieved by reader 2, and the smallest gain, 0%, would be achieved by reader 4.
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| DISCUSSION |
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Because decision making is a complex process, the simple approach of using the highest malignant rating or the average rating from multiple views may not be the method preferred by radiologists. The separate ratings that we used in this study would provide less biased information. Further investigation is needed to determine the best approach of presenting the computer's ratings to radiologists in clinical practice.
To obtain insight into how the radiologists might use the two-view information, we compared the classification results from their true two-view reading with those from a simulated two-view reading without the computer aid. The latter results were derived from ratings of single-view readings of the same 76 pairs of mammograms interpreted in experiment 2 by assuming two strategiesone in which the highest malignancy rating between the two ratings was used, and the other in which the average of the two ratings was used (Table 2). The Az values for these classification ratings derived from the single-view reading are listed in Table 2. The corresponding Az values for the computer classifier are also given in Table 2 for comparison.
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In this study, the discriminant scores of the masses given by the computer classifier were transformed into a relative malignancy rating. The relative malignancy rating scale and the distribution of the malignant and benign masses along the relative rating scale were explained to the observers in the training sessions. A relative malignancy rating scale was used because the true likelihood of malignancy of the masses could not be estimated from a small data set, as will be explained. However, the relative rating scale provided by the computer was adequate for measuring the relative performance of classification with and without CAD in an ROC study.
If a computer classifier is trained and tested with very large data sets, and if both the malignant and benign cases represent random samples of the population, then the likelihood of malignancy of a classified mass can be estimated on the basis of the probability distributions of the classifier's test output scores and the prevalence of the two classes of masses in the patient population. However, with a relatively small data set, such as that used in this and other observer studies (14), there are limitations. First, the performance of a classifier trained with a small sample set may have large bias and variance (2931). Second, the data set in this study did not include masses on which biopsy was not performed, so it did not represent a random sample of the masses in the patient population. If our classifier were applied to all cases of solid masses in clinical practice, the probability distribution of the test scores for the two classes of masses would be different from that of the current data set.
If we ignore the patient population at large, it is possible to estimate the likelihood of malignancy of a mass on the basis of the probability distribution of the classifier output scores by using the prevalence of the two classes of masses in this specific data set. However, the likelihood of malignancy derived in this way will be completely different from the true likelihood of malignancy of a mass in the patient population. This can be easily seen if one considers that the same mass with the same discriminant score will have a smaller likelihood of malignancy if it is analyzed within a data set that has a lower prevalence of malignant cases than that in the current data set.
Training the participating radiologists with a "likelihood of malignancy" derived from a small data set for the observer experiment may mislead them if they encounter a similar mass in their clinical practice. We, therefore, preferred to use a "relative malignancy rating," which is independent of the prevalences of malignant and benign masses in the data set. As long as the same classifier and the same linear transformation are used for classifying masses, the relative malignancy rating for a given mass will remain the same, regardless of the types of other masses in the data set. When a computer classifier is implemented in a clinical setting and its performance can be established in the patient population, the true likelihood of malignancy of a given mass can be estimated and provided to the radiologist. The true likelihood of malignancy may be a more informative measure for radiologists in the clinical application of CAD.
For the reading of the 76 two-view mammograms, the results of the ROC study indicated an improvement in the Az value for all six radiologists when the computer aid was used. This indicates an overall increase in the separation of confidence rating distributions between the malignant and benign cases. The histograms in Figure 10 illustrate the distributions of confidence ratings with and without CAD for reader 5, who achieved the second greatest improvement in both the Az value (Table 1) and the separation of malignant from benign distributions. Without CAD, this reader's ratings of the malignant cases ranged from 2 to 10. This is consistent with the fact that biopsy was performed in all masses in the data set to avoid missing the malignant cases. With CAD, there was marked improvement in the separation of the two distributions. It is possible to set a decision threshold at a confidence rating of 4, below which biopsy would not need to be performed and no malignant masses would be missed. The number of benign masses that could be identified without missing a malignant mass by setting an appropriate threshold would increase by 23 (out of 76 cases) for reader 5. Five of the six radiologists in our ROC study achieved an improvement in distinguishing benign from malignant masses, and one radiologist had no difference. Although the improvement of the five radiologists varied over a wide range, from one to 25 cases, this result indicates a strong possibility that CAD can be used to reduce the number of unnecessary biopsies.
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If a radiologist can increase his or her confidence in the performance of a computer aid by gaining more extensive clinical experience, then he or she will likely be able to find the most effective way of merging his or her judgment with the computer's rating and thus reduce both interobserver and intraobserver variability. Because a radiologist who uses CAD can establish a meaningful decision threshold for biopsy only after becoming familiar with the sensitivity and specificity of working with CAD, the radiologists in this study were not asked to decide whether biopsy should have been performed on a mass. Rather, we focused on the evaluation of changes in the sensitivity and specificity of the radiologists' classification of masses when CAD was used.
In this ROC study, all six observers were attending radiologists with extensive experience in the interpretation of mammograms. It is possible that the computer aid may be even more useful to radiology residents or radiologists with less experience in mammography. The effect of CAD on mammographic interpretation by less-experienced readers will be a subject of investigation in future studies.
The observers were allowed unlimited time to read each case in this ROC study. To obtain an estimate of the change in reading time with CAD, we recorded the reading time of each observer in each reading session by using a stopwatch. For the single-view reading experiment, the average reading time per image without CAD varied from 4.3 seconds to 17.1 seconds (mean time for the six observers, 7.8 seconds). The average reading time per image with CAD varied from 4.2 seconds to 17.3 seconds (mean time, 7.3 seconds). For the two-view reading experiment, the average reading time per pair of images without CAD varied from 6.6 seconds to 16.0 seconds (mean time, 10.4 seconds). The average reading time per pair of images with CAD varied from 7.6 seconds to 27.1 seconds (mean time, 13.5 seconds).
The reading time essentially did not change with use of the computer aid for the single-view readings. For the two-view readings, the radiologists took longer with CAD, probably because they had to merge the two computer ratings and merge the computer ratings with their own evaluations. Further investigation is needed to determine whether there is a trade-off between the radiologist's efficiency and the method of presenting the computer rating and whether the reading time with CAD will depend on the experience that the radiologist has with the computer information.
In the observer study, we used laser-printed mammograms instead of the original mammograms for the reading experiments. A major reason is that it is difficult to keep all the original mammograms together for the entire period of the study because they are part of active patient files and thus often recalled for comparison with new studies or for other clinical reasons. Because the maximum optical density of laser-printed images was 3.1 for the laser imager used, the contrast on the printed mammograms was about 20% lower than that on the original mammograms. Although the image quality was slightly lower than that of the original, the laser-printed digitized images were judged to be adequate for reading the details of the masses by the participating radiologists. The laser-printed image set might also be considered as one that had slightly more subtle masses than the original set of images. Because the relative performance of two modalities is measured in ROC experiments, and because the readings both with and without CAD in this study were conducted with the same set of printed images, the relative performance of the two readings should be valid. It should also be noted that in order for a computer aid that uses automated image analysis to be widely accepted, direct digital mammography would have to be the imaging modality in clinical use. Laser-printed images or soft-copy monitors will be the display medium for the digital mammograms. The use of laser-printed images for this ROC study was therefore practical.
In our observer performance experiment, we found that CAD improved the radiologists' ability to distinguish malignant and benign masses. This is consistent with the results of other studies (11,14) in which a statistically significant improvement (P < .001 in both studies) in the radiologists' classification accuracy by using CAD was found. The results of the former study (11) further showed that the PPV of a recommendation for biopsy by the radiologists was significantly increased (P < .001). In our approach, the computer classifier automatically extracted image features, whereas in the other studies, the computer classifier used the radiologist's evaluation and other patient information as input. Therefore, it appears that CAD can provide a useful second opinion to radiologists, either by consistently extracting and analyzing the image features or by optimally weighting various diagnostic factors and thereby improving the consistency in the decision-making process. This suggests that a computer classifier that combines both approachesthat is, automatically extracts image features and optimally merges them with the radiologist's evaluation and patient informationmay be even more effective for breast cancer diagnosis. The latter step will also improve the radiologist's utilization of the computer rating on the basis of the computer-extracted features; this utilization was found to have large interobserver variation in our ROC experiment.
In conclusion, an ROC study of the effects of CAD on radiologists' classification of malignant and benign masses on mammograms was conducted. The results showed that CAD can provide a statistically significant improvement in the classification accuracythat is, in the Az valuefor both single-view reading (P = .022) and two-view reading (P = .007). The improved separation between the confidence ratings of the malignant masses and those of the benign masses indicates the potential that CAD may reduce the rate of biopsy of benign masses when decision thresholds are properly chosen by the radiologists. The decision threshold may vary among radiologists, as in the case of mammographic interpretation without CAD, and can be set after the radiologist working with CAD has established his or her sensitivity and specificity with this approach through clinical experience.
Further studies are needed to evaluate the effects of CAD on the accuracy of radiologist classification of masses in clinical settings in which the prevalence of malignant masses is different from that in a laboratory data set and the likelihood of malignancy of a mass can be estimated by the computer classifier. In the two-view reading ROC experiment, the reading time per case increased by about 30% with the use of CAD. The dependence of the radiologist's efficiency in reading with CAD on the presentation method and on the reader's experience in using the computer information also warrants further investigation.
| Acknowledgments |
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| Footnotes |
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Abbreviations: CAD = computer-aided diagnosis PPV = positive predictive value ROC = receiver operating characteristic
Author contributions: Guarantor of integrity of entire study, H.P.C.; study concepts and design, H.P.C., M.A.H., B.S., N.P.; literature research, H.P.C., M.A.H.; experimental studies, M.A.H., M.A.R., T.E.W., D.D.A., C.P., J.S.N.; data acquisition, all authors; data analysis, H.P.C., B.S., N.P.; statistical analysis, H.P.C.; manuscript preparation, editing, and review, H.P.C., B.S., M.A.H., N.P., M.A.R., T.E.W., D.D.A., C.P., J.S.N.
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