Reflex UroVysion testing in suspicious urine cytology cases

BACKGROUND:

UroVysion is a US Food and Drug Administration–approved fluorescence in situ hybridization (FISH) probe set for use in the detection of recurrent urothelial carcinoma and in patients with hematuria. The objective of the current study was to evaluate the usefulness of UroVysion as a reflex test in patients with a suspicious urine cytology diagnosis. The rationale was that a more aggressive workup might be indicated in patients with a suspicious cytology diagnosis and positive UroVysion test.

METHODS:

The study population included 161 urine specimens diagnosed as suspicious over a period of 12 months. The sensitivity, specificity, negative predictive value (NPV), and positive predictive value (NPV) were calculated based on the histologic and cystoscopic correlation.

RESULTS:

The results using the reporting criteria suggested by the manufacturer demonstrated a sensitivity of 68.3%, a specificity of 39.7%, a PPV of 56.8%, and a NPV of 51.9%. The results using the presence of any cytogenetic abnormality as a positive FISH test demonstrated a sensitivity of 82.9%, a specificity of 21.7%, a PPV of 54.8%, and an NPV of 51.7%.

CONCLUSIONS:

A negative UroVysion test did not rule out the presence of low‐grade or high‐grade urothelial carcinoma in urine specimens diagnosed as suspicious. The use of less strict criteria dramatically increased the sensitivity of UroVysion FISH; however, there was a marked decrease in specificity noted. The results in this current study appear to indicate that a more aggressive workup of patients with a suspicious cytology, positive UroVysion result, and negative cystoscopic evaluation is not currently justified. Cancer (Cancer Cytopathol) 2009. © 2009 American Cancer Society.     

Evaluation of chromosomal aberrations in patients with benign conditions and reactive changes in urinary cytology

BACKGROUND:

Fluorescence in situ hybridization (FISH) is routinely used to help clarify atypical urinary cytology. However, to the authors' knowledge, little is known regarding the frequency of chromosomal aberrations in non‐neoplastic conditions that could potentially lead to false‐positive FISH results. The objective of the current study was to evaluate the frequency of chromosomal aberrations in benign cells of the urinary tract using the UroVysion FISH test.

METHODS:

The authors analyzed 77 Papanicolaou‐stained benign urine cytology specimens with reactive epithelial atypia using a FISH assay detecting the chromosomes 3, 7, and 17 and the gene locus 9p21. A positive test result was defined as an increased copy number of at least 2 chromosomes in ≥ 4 of 25 cells, or > 10 cells with a tetraploid or octaploid pattern, or homozygous or heterozygous deletion of 9p21 (≥ 12cells).

RESULTS:

FISH was positive in 27 of 77 bladder washings (35.1%) including 25 of 65 bladder washings (40.5%) and 2 of 15 voided urines (13.5%) from patients with irritative bladder (15 of 36 patients), a history of radiotherapy (7 of 12 patients), nonspecific cystitis (3 of 11 patients), hematuria (3 of 8 patients), and lithiasis (1 of 4 patients) . In 7 of 27 FISH‐positive urothelial specimens, the positivity was solely due a polyploid pattern (tetraploid/octaploid pattern) in > 10 of the cells.

CONCLUSIONS:

Chromosomal aberrations can occur in reactive urothelial cells, with a tetraploid pattern being the most common. Even an aneuploid pattern of FISH signals does not always prove malignancy because it rarely occurs in reactive urothelial cells. Correlation of FISH results with cytomorphology and patient history is crucial to avoid false‐positive diagnoses. Cancer (Cancer Cytopathol) 2011;. © 2011 American Cancer Society.     

Comparison of ImmunoCyt,UroVysion, and urine cytology in detection of recurrent urothelial carcinoma

BACKGROUND:

ImmunoCyt (uCyt) and UroVysion are ancillary studies that may aid in the detection of urothelial carcinoma in urine specimens. We compared ImmunoCyt and UroVysion to urine cytology in the ability to detect recurrent urothelial carcinoma.

METHODS:

Single voided urine samples were obtained from 100 patients who had a previous history of bladder cancer. All patients underwent cystoscopy immediately after urine sample collection. Forty‐one cystoscopically suspicious lesions were biopsied. Urine samples were divided and processed blindly and independently in 3 different laboratories for ImmunoCyt, UroVysion, and urine cytology (ThinPrep method).

RESULTS:

Of the 41 cystoscopically positive cases, most cystoscopy findings showed multiple tumors that were papillary and less than 1 cm. Biopsies showed many low‐grade tumors (54%). Overall sensitivity of cytology for low‐ and high‐grade urothelial cell carcinoma was 15% and 27%, whereas ImmunoCyt was 62% and 91% and UroVysion was 8% and 18%, respectively. Overall specificity of cytology was 97%, whereas ImmunoCyt was 63% and UroVysion was 90%.

CONCLUSIONS:

ImmunoCyt is more sensitive than either cytology or UroVysion in detecting low‐grade tumors. Both cytology and UroVysion have comparable specificity in cystoscopically negative cases. Whereas ImmunoCyt may improve the cytological detection of recurrent bladder cancer, UroVysion may be used as a confirmatory test for either cytology or ImmunoCyt. Cancer (Cancer Cytopathol) 2009. © 2009 American Cancer Society.     

A multicolor fluorescence in situ hybridization assay: A monitoring tool in the surveillance of patients with a history of non-muscle-invasive urothelial cell carcinoma: A prospective study

BACKGROUND:

Non–muscle‐invasive urothelial cell carcinoma (NMIUCC) has a high tendency to recur and affected patients must be monitored regularly using invasive cystoscopies. The aim of the current study was to compare a multicolor fluorescence in situ hybridization (FISH) assay (UroVysion) with routine follow‐up (cystoscopy and cytology) in the monitoring of patients with a previous history of NMIUCC.

METHODS:

An unselected cohort of patients under surveillance for a previous history of NMIUCC was prospectively studied. A total of 248 examinations in 223 patients were analyzed. Each exploration was comprised of cytological and FISH microscopic examination of voided urine samples and cystoscopy. The sensitivity, specificity, and positive (PPV) and negative (NPV) predictive values for tumor recurrence of all 3 techniques were determined.

RESULTS:

The sensitivities of FISH and cystoscopy were not found to be significantly different (92.9% and 82.1%, respectively). The specificities of FISH and cystoscopy were 92.7% and 89.7%, respectively. The PPV and NPV of FISH were 53.5% and 97.2%, respectively, whereas those of cystoscopy were 63.4% and 98.9%, respectively. No significant differences were found between these 2 tests. In contrast, the sensitivity and specificity of cytology were 14.3% and 99.5%, respectively.

CONCLUSIONS:

Given the lack of statistically significant differences with regard to FISH and cystoscopy results, the authors propose that FISH could be a useful monitoring tool in the surveillance of patients with a previous history of NMIUCC. Cancer (Cancer Cytopathol) 2011;. © 2011 American Cancer Society.     

Combined morphologic and fluorescence in situ hybridization analysis of voided urine samples for the detection and follow‐up of bladder cancer in patients with benign urine cytology

BACKGROUND.

Bladder cancer is among the 5 most common malignancies worldwide. Patients with bladder cancer are closely followed with periodic cystoscopies and urine cytology analyses due to the significant risk of tumor recurrence. The UroVysion fluorescence in situ hybridization (FISH) test demonstrated higher sensitivity over urine cytology in detecting bladder cancer by most comparative studies.

METHODS.

In the current study, the diagnostic usefulness of a combined cytology and FISH analysis approach was tested using the Duet automatic scanning system in patients with benign urine cytology who were being monitored for recurrent urothelial carcinoma or being assessed for various urologic symptoms.

RESULTS.

By combining the benefits of conventional cytology with molecular diagnostics, a more sensitive detection of bladder cancer was attained. All patients who had positive cystoscopy concomitantly with urine sampling were detected by combined analysis. Additional patients that developed transitional cell carcinoma during a follow‐up period of 24 months had a previous positive result on combined analysis. Only 2 patients with a negative combined analysis result presented with late disease recurrence (20 months and 22 months, respectively, after the negative test). Therefore, negative combined analysis was found to be predictive of a lack of disease recurrence for at least 12 months. In this timeframe, the overall sensitivity, specificity, negative predictive value (NPV), and positive predictive values of the combined analysis test were 100%, 65%, 100%, and 44%, respectively.

CONCLUSIONS.

Given the absolute sensitivity and NPV of the combined analysis test, the management of patients with a negative combined analysis result might be revised and allow for more flexible assessment and management of bladder cancer patients relying more on urine bound tests. Cancer (Cancer Cytopathol) 2007. © 2007 American Cancer Society.     

Digitized microscopy in the diagnosis of bladder cancer: analysis of >3000 cases during a 7-month period

BACKGROUND:

Fluorescent in situ hybridization (FISH) analysis of urine samples has proven to be a valuable adjunctive test to urine cytology for both diagnosis and monitoring recurrence of urothelial carcinoma. Automated FISH analysis has the potential to improve laboratory efficiency and to reduce interobserver and intraobserver variability, resulting in more accurate, reproducible, assay performance.

METHODS:

A total of 3200 slides containing urine specimens, hybridized with the UroVysion Bladder Cancer Kit (Abbott Molecular, Des Plaines, Illinois), a 4‐probe set for chromosomes 3, 7, 17, and 9p21, was evaluated at Acupath Laboratories. The slides were analyzed over a 7‐month period, using the Ikoniscope ‐ oncoFISH bladder Test System (Ikonisys, New Haven, Connecticut).

RESULTS:

Analysis included the incorporation of a “flagging” system developed by Acupath Laboratories to identify cases, based on specific criteria, likely to benefit from further manual review. By using US Food and Drug Administration (FDA)‐cleared scoring criteria, 96.3% of the slides could be reported directly from the automated scan, requiring no manual review of the slide. For the remaining 3.7% of the samples (all of which were very hypocellular), a manual review of each slide subsequently allowed diagnoses to be successfully reported. The average scan time was 31.7 minutes, and the average slide scan review time was 8.3 minutes.

CONCLUSIONS:

This study demonstrated the value of an automated approach to the analysis of FISH slides, affording the benefit of high‐throughput while providing the user with the necessary images and tools to quickly and accurately report a case. Cancer (Cancer Cytopathol) 2011;. © 2011 American Cancer Society.     

“FISHing” to detect urinary and other cancers

BACKGROUND:

The UroVysion Bladder Cancer Kit detects amplifications of chromosomes 3, 7, and 17, and the deletion of 9p21, by fluorescence in situ hybridization (FISH). Because manual interpretation of UroVysion FISH is time consuming and can be challenged by variable probe signal strengths and background labeling, the authors investigated an automated image analysis system to improve throughput, productivity, quality control, and accuracy.

METHODS:

The authors evaluated the interactive BioView Duet imaging system as an aid to UroVysion FISH interpretation in a 2‐armed, blinded comparison with manual screens of the same 135 consecutive cases. Manual and Duet‐assisted interpretations were compared with respect to concordance, reproducibility, and timing.

RESULTS:

Eighty‐one cases were interpreted as positive or negative with 94% concordance and a kappa value of 0.84 between manual and Duet‐aided interpretations. Three cases that ultimately were judged positive were detected with the aid of Duet but were missed with a manual screen. A final interpretation could not be given for≈25% of Duet‐scanned cases. Duet‐aided interpretation was highly reproducible for patient and control slides. Pathologist evaluation time per case was 4 minutes compared with 30 minutes for manual interpretation. Cytotechnologist involvement added 18 minutes for a total of 22 minutes, a savings of 8 minutes per case.

CONCLUSIONS:

Duet‐aided interpretations were at least equivalent to manual interpretations. The system permitted interactive review of abnormal cells and had the ability to evaluate the same cells for brightfield cytology followed by FISH. The image processing and analysis tools of the Duet system enhanced the morphology skills of cytology professionals in providing accurate interpretations. Cancer (Cancer Cytopathol) 2010. © 2010 American Cancer Society.     

Lower HER‐2/chromosome enumeration probe 17 ratio in cytologic HER‐2 fluorescence in situ hybridization for breast cancers

BACKGROUND

Fluorescence in situ hybridization (FISH) is the gold standard for assessing HER‐2 status for breast cancers, and paraffin‐embedded tissue sections are used routinely for HER‐2 FISH. Cytologic samples also are used, but to the authors' knowledge, little is known regarding the reliability of these samples. The objective of this study was to elucidate the usefulness of cytologic specimens for HER‐2 FISH testing.

METHODS

Histologic and cytologic specimens from 32 patients with invasive ductal carcinoma of the breast were subjected to comparative analysis of HER‐2 status by FISH. FISH was performed by using a PathVysion HER‐2 DNA Probe Kit according the manufacturer's instructions. The signal ratios of chromosome enumeration probe 17 (CEP17) and HER‐2 were estimated and compared. In 15 cytologic specimens, the distance between signals (HER‐2 and CEP17) and the nearest nuclear membrane were measured by using 3‐dimensional image analysis and confocal microscopy.

RESULTS

Cytologic and histologic FISH results were compared. Signal ratios of HER‐2/CEP17 were lower in cytologic specimens from 26 of 32 patients compared with the histologic material. Three‐dimensional image analysis demonstrated that the distance between the CEP17 signal and the nuclear membrane was shorter than the distance between the HER‐2 gene and the nuclear membrane.

CONCLUSIONS

The current results demonstrated that CEP17 could be lost more easily through histologic sectioning compared with the cytology results, because CEP17 is closer to the nuclear membrane. FISH analysis in cytologic specimens produced more accurate HER‐2/CEP17 ratios, because the whole nucleus was subjected to FISH testing, compared with matched histologic specimens. Cancer (Cancer Cytopathol) 2008. © 2008 American Cancer Society.     

Value of p16(INK4a) in the diagnosis of low-grade urothelial carcinoma of the urinary bladder in urinary cytology

BACKGROUND:

The sensitivity of urinary cytology for the diagnosis of urothelial carcinomas is low, particularly in low‐grade carcinomas. The UroVysion test is a fluorescent in situ hybridization multiprobe assay that increases the sensitivity of urinary cytology. However, this test is not widely available. P16^INK4a, a protein involved in cell cycle progression, is overexpressed in urothelial carcinoma. Immunocytochemical expression of p16^INK4a has been examined in biopsy samples from urothelial carcinomas, but few studies have addressed this protein in urine cytology.

METHODS:

The authors compared the results of p16^INK4a immunoreactivity in cytology and biopsy samples from 83 cases, including low‐grade urothelial carcinomas, reactive epithelial lesions, and negative cases.

RESULTS:

p16^INK4a assessment of in urine cytology samples showed a sensitivity of 66.7% and a specificity of 82.8% in the diagnosis of low‐grade urothelial carcinomas.

CONCLUSIONS:

On the basis of these results, the authors propose that immunocytochemical detection of p16^INK4a is a reliable tool in urine cytology, both for the diagnosis of low‐grade urothelial carcinomas and for follow‐up purposes. More retrospective and prospective studies are required to verify these results. Cancer (Cancer Cytopathol) 2012. © 2012 American Cancer Society.     

Equivocal cytology in lung cancer diagnosis: improvement of diagnostic accuracy using adjuvant multicolor FISH, DNA-image cytometry, and quantitative promoter hypermethylation analysis

BACKGROUND:

Sometimes, cytological lung cancer diagnosis is challenging because equivocal diagnoses are common. To enhance diagnostic accuracy, fluorescent in situ hybridization (FISH), DNA‐image cytometry, and quantitative promoter hypermethylation analysis have been proposed as adjuncts.

METHODS:

Bronchial washings and/or brushings or transbronchial fine‐needle aspiration biopsies were prospectively collected from patients who were clinically suspected of having lung carcinoma. After routine cytological diagnosis, 70 consecutive specimens, each cytologically diagnosed as negative, equivocal, or positive for cancer cells, were investigated with adjuvant methods. Suspicious areas on the smears were restained with the LAVysion multicolor FISH probe set (Abbott Molecular, Des Plaines, Illinois) or according to the Feulgen Staining Method for DNA‐image cytometry analysis. DNA was extracted from residual liquid material, and frequencies of aberrant methylation of APC, p16^INK4A, and RASSF1A gene promoters were determined with quantitative methylation‐specific polymerase chain reaction (QMSP) after bisulfite conversion. Clinical and histological follow‐up according to a reference standard, defined in advance, were available for 198 of 210 patients.

RESULTS:

In the whole cohort, cytology, FISH, DNA‐image cytometry, and QMSP achieved sensitivities of 83.7%, 78%, 79%, and 49.6%, respectively (specificities of 69.8%, 98.2%, 98.2%, and 98.4%, respectively). Subsequent to cytologically equivocal diagnoses, FISH, DNA‐image cytometry, and QMSP definitely identified malignancy in 79%, 83%, and 49%, respectively. With QMSP, 4 of 22 cancer patients with cytologically negative diagnoses were correctly identified.

CONCLUSIONS:

Thus, adjuvant FISH or DNA‐image cytometry in cytologically equivocal diagnoses improves diagnostic accuracy at comparable rates. Adjuvant QMSP in cytologically negative cases with persistent suspicion of lung cancer would enhance sensitivity. Cancer (Cancer Cytopathol) 2011;. © 2011 American Cancer Society.     

Cytogenetics and fluorescence in situ hybridization as adjuncts to cytology in the diagnosis of malignant mesothelioma

BACKGROUND:

Virtually all malignant mesotheliomas (MMs) exhibit clonal chromosomal aberrations. Although the chromosome regions affected by these aberration(s) may vary from 1 tumor to another, certain regions are commonly disrupted. These aberrations are absent in benign mesothelial cells, and therefore their presence can be used to confirm a diagnosis of MM. In the current study, the authors investigated the value of karyotyping and fluorescence in situ hybridization (FISH) as adjuncts to conventional cytologic examination in patients with MM.

METHODS:

A retrospective analysis of 48 pleural or peritoneal fluids from patients with histologically confirmed MM was performed. Karyotypic analyses were attempted in all cases. In 27 cases, FISH for deletions of 9p21 (CDKN2A gene) and 22q was also performed because the karyotype was normal or unsuccessful.

RESULTS:

Karyotypes were obtained in 35 (73%) of the specimens. Of these, 15 (43%) were abnormal and 20 (57%) were normal. Thirteen additional abnormal results were detected by FISH in cases for which the karyotypes were normal or unsuccessful. A total of 24 cases (50%) had an associated cytologic interpretation. Karyotyping or FISH was abnormal in 8 cases that were interpreted cytologically as either negative or suspicious.

CONCLUSIONS:

The combination of FISH and karyotyping was found to improve on the diagnostic sensitivity of karyotyping alone in detecting MM in effusions. The authors concluded that karyotyping and FISH together were a more useful adjunct to cytology than FISH or karyotyping alone. Cancer (Cancer Cytopathol) 2009. © 2009 American Cancer Society.     

The application of cytogenetics and fluorescence in situ hybridization to fine‐needle aspiration in the diagnosis and subclassification of renal neoplasms

BACKGROUND:

Percutaneous fine‐needle aspiration (FNA) cytology is an important diagnostic test for the evaluation and management of selected renal masses. Cytogenetic analysis of cytology specimens can serve as an adjunct for precise classification because certain tumors are associated with specific chromosomal aberrations. This study summarizes our experience with the application of conventional cytogenetics and fluorescence in situ hybridization (FISH) to renal FNA specimens.

METHODS:

All percutaneous renal FNAs performed during 2005 through 2008 were identified from the electronic pathology database. Results of cytogenetic and FISH analyses were correlated with the final diagnoses of the renal FNAs.

RESULTS:

A total of 303 renal FNAs were performed. During an onsite assessment, a portion of the cytology specimen was allocated for cytogenetic analysis in 74 cases. Karyotypic analysis or FISH was successful in 44 (59%) of these. Characteristic chromosomal abnormalities were observed in 27 cases. In 17 cases, a karyotype revealed a combination of trisomies/tetrasomies and in another 5 cases, FISH revealed trisomy 7 and 17, both of which are consistent with papillary renal cell carcinoma (RCC). Two cases showed 3p deletions consistent with clear cell RCC. Trisomy 3 was observed in 1 case of clear cell RCC. Monosomy 1 and 17 was observed in a case of papillary RCC comprised oncocytic cells. In 1 case of primary renal synovial sarcoma, FISH revealed a rearrangement at the SYT locus (18q11.2).

CONCLUSIONS:

Renal FNA specimens are amenable to analysis by cytogenetics and FISH in the diagnosis and subclassification of renal neoplasms. Cancer (Cancer Cytopathol) 2010. © 2010 American Cancer Society.     

Analysis of ThinPrep cytology in establishing the diagnosis of small cell carcinoma of lung

BACKGROUND:

ThinPrep liquid‐based cytology (TP) preparations are being used increasingly in nongynecologic specimens. To the authors' knowledge, few studies to date have evaluated TP cytology in the setting of small cell carcinoma of the lung (SCCL). Accurately differentiating SCCL from other lung tumors has important clinical and therapeutic implications. Herein, the authors evaluated the diagnostic utility and cytomorphology of TP in the setting of SCCL.

METHODS:

All cases of SCCL with prior or concurrent TP cytologic specimens were identified via computer search. The cytodiagnoses were tabulated. When available, cytologic material was reviewed. Performance parameters of the various cytologic modalities processed with TP were calculated.

RESULTS:

In 121 patients with SCCL, 261 TP specimens were identified. The cytodiagnoses were: SCCL (119 specimens), suspicious for SCCL (45 specimens), atypical cells‐not otherwise specified (22 specimens), negative/nondiagnostic (63 specimens), and nonsmall cell carcinoma (4 specimens). During the same period of time, 3 cases of false‐positive diagnoses of SCCL were identified. The positive predictive value for a cytodiagnosis of SCCL on TP was 97.5%, and the sensitivity was 62.8%. Bronchial brush was the most sensitive cytologic modality (78.3%). Immunostaining was found to be contributory to the diagnosis in 10 of the 11 cases in which it was attempted.

CONCLUSIONS:

TP is a sound alternative to conventional preparations for the cytodiagnosis of SCCL. Cytomorphology of SCCL is altered in TP with less molding, less cell fragility, more discohesion, and tumor cell shrinkage artifact. Immunohistochemical staining of cellblocks is a useful adjunct in challenging cases. A positive cytodiagnosis of SCCL on TP can be used to initiate definitive therapy when biopsy is not possible. Cancer (Cancer Cytopathol) 2009. © 2009 American Cancer Society.     

Utility of paired box gene 8 (PAX8) expression in fluid and fine‐needle aspiration cytology

BACKGROUND:

Metastases from ovarian neoplasms are commonly encountered in peritoneal fluids. In addition, reactive mesothelial cells in effusion specimens can mimic ovarian serous carcinoma, making the diagnosis difficult. Calretinin has been recognized as a reliable immunohistochemical marker for mesothelial cells, whereas WT1 has proven useful in the diagnosis of ovarian serous carcinoma. This can present a diagnostic pitfall in effusion cytology, because mesothelial cells can demonstrate immunoreactivity for WT1. Recently, paired box gene 8 (PAX8) has been used in distinguishing ovarian from mammary carcinoma. To the authors' knowledge, no studies using PAX8 have been performed on peritoneal cytology specimens to date, and its expression in metastatic ovarian serous carcinoma has not been studied.

METHODS:

These markers, along with BerEP4 and MOC‐31, were evaluated in cytology cell block preparations from 30 fluid cytology specimens and 11 fine‐needle aspiration specimens.

RESULTS:

PAX8 was found to be positive in 37 of 41 (90%) ovarian carcinoma cases studied, and was a sensitive (90%) and specific (100%) marker for the detection of metastatic ovarian carcinoma. In addition, calretinin was found to be useful for identifying mesothelial cells in fluid cytology. Furthermore, although PAX8 and WT1 have demonstrated comparable sensitivity (90% and 93%, respectively) in diagnosing metastatic ovarian carcinoma, PAX8 appears to have superior specificity because staining is not observed in mesothelial cells. BerEP4 and MOC‐31 were found to have a lower sensitivity and specificity compared with PAX8.

CONCLUSIONS:

PAX8‐positive, calretinin‐negative staining appears to be highly specific and sensitive for detecting metastatic ovarian serous carcinoma in cytologic preparations and can be useful in distinguishing it from mesothelial cells in fluid cytology. Cancer (Cancer Cytopathol) 2010. © 2010 American Cancer Society.     

Multi-target fluorescence in situ hybridization in bladder washings for prediction of recurrent bladder cancer

The objective of this study was to evaluate the diagnostic value of chromosomal analysis by fluorescence in situ hybridization (FISH) for predicting recurrence of urothelial carcinoma (UC) after transurethral resection. One hundred and thirty-eight patients (median age 68.5 years) with a history of UC were eligible for this prospective study. FISH was applied to cytospin specimens prepared from bladder washings taken during a negative control cystoscopy. The multi-target FISH test UroVysion (Abbott/Vysis) containing probes to the centromeres of chromosomes 3, 7, 17 and the 9p21 locus was used. UC recurrence was defined as a positive biopsy during follow-up. The median follow-up time was 19.2 (4-52) months. FISH was positive in 50 (36%) patients and negative in 88 (64%) patients. A recurrence occurred in 39% of the patients with a positive FISH test and in 21% of patients with a negative FISH test. FISH positivity according to manufacturer's criteria, at the time of a negative cystoscopy, was not significantly associated with the risk of recurrence (p = 0.12). However, the sensitivity of the FISH test to predict recurrence was significantly improved by considering specimens with rare (< or =10) tetraploid cells as negative (p < 0.006). In addition, presence of 9p21 deletion was significantly associated with recurrence (p < 0.01). Notably, positive standard cytology was an independent factor for subsequent recurrence in this study (p < 0.001). Taken together, multi-target FISH may help to stratify the risk of recurrence of UC at the time of a negative follow-up cystoscopy. Defining the optimal threshold for FISH positivity requires consideration of tetraploid pattern and 9p21 deletion. Our results also emphasize the paramount importance of conventional cytology for UC surveillance.     

Role of BRAFV600E mutation analysis and second cytologic review of fine-needle aspiration for evaluating thyroid nodule

BACKGROUND:

Thyroid fine‐needle aspiration (FNA) is the primary diagnostic tool used for the evaluation of thyroid nodules. Although most aspirates provide diagnostic cytology, some are classified as indeterminate. The aim of this study was to determine whether the second review of FNA cytology can improve the diagnostic values and to assess the role of proto‐oncogene B‐Raf (BRAF) mutation testing in the diagnosis of papillary carcinoma (PC).

METHODS:

Thyroid aspirates from 1060 patients were submitted for cytologic evaluation and BRAFV600E mutation analysis. A second cytologic review was performed by 2 cytopathologists in light of the mutation status.

RESULTS:

Of the 313 patients who received surgery, 200 (63.9%) had been initially diagnosed as malignant by cytology. They were surgically confirmed as PCs, and the BRAFV600E mutation was detected in 82.5% of the cases. Ninety‐five of 102 cases (93.1%) with indeterminate cytology turned out to be malignant, and the mutation was present in 63.3% of PCs. The sensitivity, accuracy, and negative predictive value (NPV) of the second review were better than those of initial cytologic diagnosis (P <.001). The addition of the mutation analysis significantly increased the sensitivity, accuracy, and NPV for detecting PCs compared with those of cytology alone.

CONCLUSIONS:

Qualified cytologic diagnosis increases the effectiveness of FNA, forgoing the need for repeat biopsy or intraoperative frozen section evaluation. Preoperative BRAF mutation testing can supplement the routine cytology in the selection of thyroid nodules for surgery. Cancer (Cancer Cytopathol) 2012. © 2011 American Cancer Society.     

Peritoneal washing cytologic analysis of ovarian serous tumors of low malignant potential to detect peritoneal implants and predict clinical outcome

BACKGROUND:

Patients with ovarian serous tumors of low malignant potential (OSLMP) who have peritoneal implants, especially invasive implants, are at an increased risk of developing tumor recurrence. To the best of the authors' knowledge, the ability of peritoneal washing (PW) cytology to detect the presence and type of peritoneal implants has not been adequately investigated, and its prognostic significance is unknown.

METHODS:

Records and PW specimens of 101 patients diagnosed with and treated for OSLMP between 1996 and 2010 at The University of Texas MD Anderson Cancer Center were retrospectively reviewed. Patients' staging biopsy findings were compared with the results of the authors' review of the PWs. Follow‐up data were also analyzed.

RESULTS:

Of the 96 patients for whom staging biopsy results were available, 26 (27%) had peritoneal implants (17 noninvasive and 9 invasive), 19 (20%) had endosalpingiosis, and 51 (53%) had negative findings. The PW specimens of 18 of the 26 patients (69%) with peritoneal implants were positive for serous neoplasm, and a correlation was found between cytologic and histologic findings (P < .0001). The sensitivity, specificity, positive predictive value, and negative predictive value were 69%, 84%, 62%, and 88%, respectively. Four of 101 patients had disease recurrence; 3 of these patients had invasive implants and 1 patient had noninvasive implants. None of the patients who had negative staging biopsy findings or endosalpingiosis but did have PW specimens that were positive for serous neoplasm developed disease recurrence.

CONCLUSIONS:

PW cytology detects the presence of peritoneal implants with moderate accuracy. However, long‐term studies are needed to determine whether positive PW cytologic findings are an independent predictor of tumor recurrence. Cancer (Cancer Cytopathol) 2012;. © 2012 American Cancer Society.     

Evaluation of quantity and staining pattern of human papillomavirus (HPV)‐infected epithelial cells in thin‐layer cervical specimens using optimized HPV‐CARD assay

BACKGROUND.

Testing for human papillomavirus (HPV) is used in the triage of women with a cervical cytology of atypical squamous cells of undetermined significance (ASCUS). A fluorescent in situ hybridization assay was developed for the detection of HPV using the catalyzed receptor deposition technique (HPV‐CARD). In this study, the utility of this assay was tested for the detection of HPV in liquid‐based cervical cytology specimens.

METHODS.

A total of 195 liquid‐based cytology specimens were analyzed using the HPV‐CARD assay. The results from the assay were compared with HPV polymerase chain reaction (PCR) and typing results. The number of HPV‐infected cells and the staining pattern was correlated with the cytology classification.

RESULTS.

A 91% concordance between HPV‐CARD and PCR was observed for the detection of high‐risk HPV viruses. A 78% concordance was observed for specimens that were negative for HPV. In ASCUS, low‐grade squamous intraepithelial lesion (LSIL), and high‐grade squamous intraepithelial lesion (HSIL) categories, the average number of HPV‐positive cells per slide was 19 cells, 127 cells, and 450 cells, respectively. The number of cells with a punctate staining, suggestive of HPV integration, was 21% in ASCUS, 34% in LSIL, and 46% in HSIL specimens.

CONCLUSIONS.

The results of the current study indicate positive correlations between the severity of the disease and the increased overall quantity of HPV‐positive epithelial cells in cervical cytology specimens and accumulation of cells with punctate staining suggestive of integrated HPV. In summary, the developed HPV‐CARD assay was found to provide novel information regarding the proportion and staining pattern of HPV‐infected epithelial cells in different cytologic categories of cervical specimens. Cancer (Cancer Cytopathol) 2007. © 2007 American Cancer Society.     

DNA image cytometry and fluorescence in situ hybridization for noninvasive detection of urothelial tumors in voided urine

BACKGROUND: Cystoscopy and histologic examination remain the standard methods for initial tumor diagnosis and monitoring for early detection of recurrences, since the sensitivity of conventional urinary cytology for the detection of urothelial tumors in urinary specimens is low. DNA image cytometry (ICM) and fluorescence in situ hybridization (FISH) have been suggested as ancillary tools. The goal of the current study was to compare the diagnostic value of DNA image cytometry and FISH for the noninvasive detection of urothelial tumors in voided urine. METHODS: Cytospin preparations were prepared from voided urine collected prior to the resection of 26 noninvasive (pTa) and 11 invasive (pT1-2) tumors. Specimens from 14 patients with benign prostatic hyperplasia were used as negative controls. DNA ICM was performed using the AUTOCYTE trade mark cell analytical system on Feulgen-stained cytospin specimens. The commercially available UroVysion trade mark FISH multiprobe was used to analyze chromosomes 3, 7, and 17, and 9p21. RESULTS: The overall sensitivity of cytology improved from 24% to 54% and to 78% if supplemented by ICM or FISH, respectively. Image cytometry detected all invasive tumors (pT1-2), while FISH missed one; FISH identified 19 of 26 (73%) pTa tumors, while only 9 (35%) of these tumors were aneuploid by ICM. The results of ICM and FISH were concordant in 37 of 51 (72%) cases. CONCLUSIONS: The current study shows that both FISH and ICM can successfully be used as supplementary methods to detect the clinically most relevant group of invasive bladder carcinomas. However, UroVysion FISH is more sensitive in the detection of pTa tumors than ICM, as it recognizes individual chromosomal alterations that frequently prevail in urothelial tumors.