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PubMed2000

A staining procedure for micronucleus test using new methylene blue and acridine orange: specimens that are supravitally stained with possible long-term storage.

The micronucleus test has been widely used as an in vivo cytogenetic test. It employs two different kinds of supravital staining methods which use either new methylene blue (N) and Giemsa (G) or acridine orange (AO). We have developed a new staining procedure for the preparation of specimens supravitally stained with possible long-term storage, using both N and AO. This N/AO-staining method involves three steps; (1) combination of the target tissue or target cells with an equivalent volume of 0.5% solution of new methylene blue (N-staining step), (2) immediate smear of the mixture, followed by treatment with methanol for 10 min for fixation and removal of N and drying (referred to as fixed-decolorized specimens), and (3) staining with 0.007% solution of AO for 3 min, followed by washing with Sörensen's buffer (pH 6.8) and covering of specimens before observation (AO-staining step). To examine whether the N/AO-staining method is useful for the micronucleus test, comparisons were made between N-, N/AO-, and AO-stained specimens prepared supravitally from peripheral blood of rats with and without treatment of cyclophosphamide. The results indicate that N/AO-stained specimens can be supravitally observed after long-term storage with the same coloration and comparable frequencies of micronucleated reticulocytes with a positive response as AO-stained specimens, if the staining process is temporarily stopped before AO-staining (as fixed-decolorized specimens), or if the AO-staining step is repeated. The results also showed that separated reticulocyte types are supravitally stained in a similar fashion to N-stained specimens but not to AO-stained specimens, indicative of the preservation of the supravital feature of N-staining. Taken together these results suggest that the N/AO-staining procedure could offer an additional useful staining tool for the micronucleus test.

Acridine Orange↗

PubMed1998

Malignant melanoma re-excision specimens--how many blocks?

AIMS: Wide local excision is commonly undertaken as part of the further management of cutaneous melanoma. Although the original excision is usually complete, pathologists vary considerably in their macroscopic handling and sampling of the wide excision specimens. This study evaluates the sampling of reexcision specimens and the information gained from histological examination of tissue blocks in order to develop guidelines for the handling of these specimens. METHODS AND RESULTS: The study group comprises 219 cases of melanoma which underwent initial biopsy followed by wider excision. The macroscopic appearance, number of blocks taken and presence of residual melanoma in the wide excision specimen was assessed. Residual melanoma was detected in the wide excision specimen in only four cases out of the 167 in which the original melanoma had been described as completely excised and in these cases the detection of a residual lesion reflected either advanced local disease at the time of the original excision, or an incompletely excised extensive radial growth phase. The sampling of wide excision specimens varied (range of blocks 1-12, average 3.1) but statistical analysis showed no undersampling of the wide excision specimens in which no residual lesion was found. CONCLUSIONS: These results show that detection of a residual lesion in wide excision specimens of melanomas previously completely excised is unusual and offers no additional clinically useful information. Random block taking in the absence of a macroscopic abnormality is therefore unlikely to be clinically useful. Guidelines for the gross handling of these wide excision specimens are proposed which are likely to save resources in most histopathology departments by reducing block numbers.

Humans↗

PubMed1987

Influence of specimen treatment on nonreactive HTLV-III sera.

We investigated effects of specimen treatment on HTLV-III antibody assay results. Freezing and thawing specimens 10 times did not increase specimen mean absorbance (A) values measured by the Abbott immunoenzymometric assay (IEMA). Heating blood donor serum specimens at 56 degrees C for at least 10 min inactivates virus, but converted 7.1% of nonreactive specimens to positive when measured by this IEMA. Also heating at 56 degrees C for 30 min increased A values of most specimens; however, the conversion to positive within assay ranged from 0 to 75%, with a mean of 17%. When 56 nonreactive patient specimens were heated, 55% became Abbott IEMA reactive, although ENI IEMA values did not increase and Western blot results remained nonreactive. Binding of immunoglobulins G and A to Abbott IEMA beads did not cause this conversion after heat treatment. We postulate that heating specimens causes alteration of some serum matrices and introduces nonspecific binding to assay bead antigen or antibody reagents. We recommend that for the Abbott IEMA, specimens may be stored frozen, and must not be heated to avoid altered reactivity.

Antibodies, Viral↗

PubMed1996

Lymph node recovery from colorectal resection specimens removed for adenocarcinoma. Trends over time and a recommendation for a minimum number of lymph nodes to be recovered.

Recovery of pericolorectal lymph nodes from colectomy specimens has long been part of colorectal cancer staging. Recently, adjuvant therapy has been added for high stage carcinomas, providing further impetus for performing careful lymph node dissections. Pericolorectal lymph nodes were examined to determine if there has been a change over time in the number of lymph nodes recovered and proportion of specimens with pericolonic lymph node metastases from colorectal carcinoma resection specimens. Also, the authors attempted to establish a recommendation for a minimum number of lymph nodes that should be recovered before a colon can be considered free of metastases. Slides and reports of the first 20 consecutive pT3 colorectal carcinoma resections in each year from 1955 to 1995 at William Beaumont Hospital that did not have known metastases at the time of surgery were reviewed (750 specimens total). The mean number of lymph nodes recovered per specimen and incidence of detected lymph node metastases increased over the 41-year period, with the greatest increase occurring during 1992-1995. The greatest proportion of patients with lymph node metastases detected occurred in the 17 to 20 lymph nodes recovered per specimen group. Specimens with more than 20 lymph nodes did not have a higher proportion of lymph node metastases detected compared to specimens with 17 to 20 lymph nodes. Approximately 20% of the specimens with metastases had more than 17 lymph nodes recovered. These results suggest that pathologists should retrieve all the lymph nodes that can be recovered, but at least 17 lymph nodes should be recovered to insure accurate documentation of nodal metastases when present.

Adenocarcinoma↗

PubMed1999

Detection of Chlamydia trachomatis and Neisseria gonorrhoeae in swab specimens by the Hybrid Capture II and PACE 2 nucleic acid probe tests.

BACKGROUND AND OBJECTIVES: The Digene Hybrid Capture II (HC II) CT/GC Test (Digene Corp., Beltsville, MD) is a new nucleic acid signal amplification-based test for the detection of Chlamydia trachomatis and Neisseria gonorrhoeae in specimens from the genital tract. For optimal results, the HC II CT/GC Test employs a special conical shaped brush for cervical specimen collection from nonpregnant women and swabs from pregnant women. GOALS: To validate a protocol for HC II C. trachomatis and N. gonorrhoeae testing of specimens collected for the GenProbe PACE 2 System. STUDY DESIGN: Specimens were collected from 1,746 patients with a swab and placed in GenProbe transport media according to the manufacturer's recommended procedure. The specimens were first tested at two clinical laboratories by the PACE 2 system, and then blindly tested by HC II CT/GC using an adjusted cutoff value. Discrepant specimens were adjudicated by polymerase chain reaction (PCR), and the result common to two of the three testing methods (HC II, PACE 2, and PCR) was defined as the consensus result. RESULTS: Combining the data from both sites, the relative sensitivity of the HC II Test compared with the consensus result for the detection of 1,761 specimens for C. trachomatis and 1,750 specimens for N. gonorrhoeae was 100% for both organisms. The relative specificities for C. trachomatis and N. gonorrhoeae detection were 99.8% and 99.7%, respectively. The relative sensitivities of the PACE 2 CT and GC Systems were 86.5% and 87.1%, respectively, with relative specificities of 99.9% and 100%. The difference in sensitivity between HC II and PACE 2 for C. trachomatis detection was significant (P < 0.016). CONCLUSION: The HC II CT/GC Test can be performed using specimens collected in GenProbe transport media and has a significantly greater sensitivity for C. trachomatis detection than the PACE 2 System.

Cervix Uteri↗

PubMed1995

Evaluation of nucleic acid-based test (PACE 2C) for simultaneous detection of Chlamydia trachomatis and Neisseria gonorrhoeae in endocervical specimens.

A nucleic acid-based test (Gen-Probe PACE 2C System) was evaluated for the ability to detect Chlamydia trachomatis and Neisseria gonorrhoeae from endocervical specimens in a single assay. Three swab samples, randomized for collection order, were obtained from each patient and tested by N. gonorrhoeae and C. trachomatis culture and by the PACE 2C probe assay. Fifty of 395 specimens were culture positive for N. gonorrhoeae (17 specimens), C. trachomatis (26 specimens), or both (7 specimens), of which PACE 2C testing detected 48 specimens. The PACE 2C assay was positive for 56 specimens, including 8 specimens not positive by culture. Of the total of 10 discrepancies between culture and PACE 2C results, resolution testing yielded four false-negative culture, four false-positive PACE 2C, and two false-negative PACE 2C results. The sensitivity, specificity, and positive and negative predictive values for PACE 2C after reevaluation were 96.3, 98.8, 92.9 and 99.4%, respectively. The overall sensitivities for C. trachomatis and N. gonorrhoeae culture were 89.2 and 88.9%, respectively. The prevalence rate for C. trachomatis was 9.4%, and that for N. gonorrhoeae was 6.8%. The Gen-Probe PACE 2C System is a reliable alternative for screening endocervical specimens for both C. trachomatis and N. gonorrhoeae in a single assay.

Cervix Uteri↗

PubMed2000

Relationship of incremental specimen volumes and enhanced detection of human immunodeficiency virus type 1 RNA with nucleic acid amplification technology.

The relationship between specimen input volume and the frequency of reported human immunodeficiency virus type 1 (HIV-1) RNA copy numbers by nucleic acid amplification technology (the NASBA HIV-1 RNA QT system) was investigated. Results obtained with both clinical specimens and dilution panels indicated that both the absolute number of reported results and the reported HIV-1 RNA copy number were directly proportional to the specimen input volumes evaluated (0.1, 0.5, and 1.0 ml). Conversion of the reported HIV-1 RNA copy numbers to a constant 1.0-ml volume indicated that the numerical relationship among the specimen input volumes and the HIV-1 RNA copy numbers was multiplicative. The HIV-1 RNA copy numbers reported for the 0.5-ml input volume were approximately 5-fold increased over those reported for the 0.1-ml input volume, and those reported for the 1.0-ml input volume were 10-fold increased over those reported for the 0.1-ml input volume. For the specimen input volumes investigated, a common linear range of 264 to 5,400,000 HIV-1 RNA copies was observed. The use of increased specimen input volumes did not result in a loss of assay specificity, as the results reported for specimens from 50 seronegative blood donors were negative at all three specimen input volumes. In conclusion, an increase in the input volume of specimens analyzed by nucleic acid amplification technology can be useful for the enhanced detection of HIV-1 RNA.

CD4 Lymphocyte Count↗

PubMed2005

Evaluation of PCR testing of ethanol-fixed nasal swab specimens as an augmented surveillance strategy for influenza virus and adenovirus identification.

Viral culture isolation has been widely accepted as the "gold standard" for laboratory confirmation of viral infection; however, it requires ultralow temperature specimen storage. Storage of specimens in ethanol at room temperature could expand our ability to conduct active surveillance and retrospective screenings of viruses with rapid and inexpensive real-time PCR tests, including isolates from remote regions where freezing specimens for culture is not feasible. Molecular methods allow for rapid identification of viral pathogens without the need to maintain viability. We hypothesized that ethanol, while inactivating viruses, can preserve DNA and RNA for PCR-based methods. To evaluate the use of ethanol-stored specimens for augmenting surveillance for detection of influenza viruses A and B and adenoviruses (AdV), paired nasal swab specimens were collected from 384 recruits with febrile respiratory illness at Fort Jackson, S.C., in a 2-year study. One swab was stored at ambient temperature in 100% ethanol for up to 6 months, and the other swab was stored at -70 degrees C in viral medium. For viral detection, frozen specimens were cultured for a variety of respiratory viruses, and ethanol-fixed specimens were tested with TaqMan (TM) probe and LightCycler SYBR green (SG) melting curve assays with at least two different PCR targets for each virus. The sensitivities of the TM and SG assays on specimens stored in ethanol for 1 month were 75% and 58% for influenza A, 89% and 67% for influenza B, and 93 to 98% and 57% for AdV, respectively. Lower specificities of the real-time assays corresponded to the increased detection of PCR-positive but culture-negative specimens. Influenza virus RNA was detected as well or better after 6 months of storage in ethanol.

Adenovirus Infections, Human↗

PubMed1996

Microcalcifications in breast core biopsy specimens: disappearance at radiography after storage in formaldehyde.

PURPOSE: To determine whether radiographically proved calcifications in core biopsy specimens are better preserved in a nonaqueous fixative than in aqueous solutions. MATERIALS AND METHODS: One hundred fifty core biopsy specimens were taken from two female breast specimens that had been preserved in ethanol. Forty-one of the specimens with radiographically proved microcalcifications were divided into four groups and deposited in four different solutions: 10% formaldehyde, 0.9% sodium chloride, electrolyte solution, and 74.1% ethanol with 10% 2-propanol. The core specimens were radiographed again after 1 and 3 days. Five ethanol-preserved specimens were also reexamined radiographically after 2 weeks. RESULTS: Within 3 days, total radiographic disappearance of microcalcifications was observed in all core biopsy specimens that were immersed in solutions with high water content. In those core specimens preserved in ethanol, microcalcifications showed no change. CONCLUSION: A nonaqueous fixative, such as ethanol, is a better preservative of microcalcifications in breast core biopsy specimens than various aqueous solutions, possibly because calcium compounds are water-soluble.

Biopsy↗

PubMed1999

Surgical pathology specimens for gross examination only and exempt from submission: a College of American Pathologists Q-Probes study of current policies in 413 institutions.

OBJECTIVE: To survey the scope of current written institutional policies for types of surgical pathology specimens exempt from submission to the laboratory and those that may be examined by gross inspection only. DESIGN AND SETTING: In the first quarter of 1997, a total of 413 voluntary participant institutions enrolled in the College of American Pathologists Q-Probes quality improvement program completed a checklist of 115 proffered specimens. Also included was a questionnaire defining demographic, practice, and reimbursement variables. MAIN OUTCOME MEASURES: The number and types of specimens exempt from submission and submitted for gross examination only based on written institutional policy rather than on unapproved actual practices. RESULTS: Most institutions had a written policy for types of specimens deemed exempt from submission to pathology (87.1 %) and for types of specimens subject to gross examination only (76.6%). There was a wide range of numbers of specimen types with a median number of 17 (range, 2-40) in the exempt category and 29 (range, 6-57) in the category of gross examination only. Significantly higher absolute counts of specimens exempt from submission to pathology were reported by institutions with a greater surgical pathology volume in 1996 and by nonteaching institutions. No aspect of practice was associated with numbers of specimens for gross examination only. CONCLUSIONS: This Q-Probes study creates a multi-institutional reference database of current practices to assist pathologists and clinical staff in the development of written guidelines pertaining to surgical pathology specimen submission exceptions and gross-only examinations.

Biopsy↗

PubMed2005

[A project to reduce mistakes in collecting specimens].

Specimen collection is an important task of nursing care. Any failure to collect the whole missing of a specimen may affect diagnosis and treatment. It will also increase the workloads of staff and the cost of medicines. From April to July 2002, a total of 3,382 specimens were collected in A112 ward in Taipei Veterans General hospital. There were 43 wrong specimens, 1.27% of the total. Analysis showed that the reasons for the error were as follows: (1) The level of knowledge about the test tubes and about the volume of specimens was low. (2) The procedure for dealing with application from was not appropriate. (3) The box in which specimens were kept was not appropriate. (4) The checklist was bad. The PDCA process used in this project reduced the rate of wrong specimens. The strategy was designed with three dimensions: staff, facilities, and procedure. After it was implemented, the rate of wrong specimens dropped from 1.27% to 0.49%, the improvement rate was 61.4%, and the rate of achievement was 101.3%. This project not only reduced the rate of wrong specimens, but also made the process easier and more safisfying for staff.

Humans↗

PubMed2004

Native Hawaiian preferences for informed consent and disclosure of results from research using stored biological specimens.

Increasingly, genetic and biomedical researchers are developing protocols to reexamine human tissue specimens that were obtained and stored during clinical care or previous research studies. Although some communities and associations are developing guidelines for human-tissue research, guideline development rarely considers consumer preferences for informed consent and disclosure of results. This study, examining Native Hawaiian preferences for informed consent and disclosure of results (n = 429, 83.2% Native Hawaiian), was modeled after a national study of consumer preferences, allowing comparison between the national sample and the Hawai'i-based sample. The interview schedule included two scenarios on research requiring the re-use of clinically derived and research-derived biological specimens. For each, participants were asked if informed consent should be required: a) in general; b) if the specimen was personally identified; and c) if the specimen was de-identified, or anonymized. Participants were also asked if they would want to know the results of the research and if they would want their doctor to be told. Regardless of how specimens were obtained, 78% of Native Hawaiians would want to be asked for their consent for the re-use of identified specimens and about 35% would want to be consented for the re-use of anonymized specimens. In both cases, Native Hawaiians in the Hawai'i sample were more likely than Whites in the national sample to want an informed consent process. Similar proportions in both samples would want findings from research on stored specimens reported to them (about 90%) and to their physicians (about 80%). These findings call into question the "Common Rule" and the guidelines of the American Society of Human Genetics, which do not require researchers to obtain informed consent for research use of anonymized specimens.

Adolescent↗

PubMed1987

Microbiologic evaluation of canine urine: direct microscopic examination and preservation of specimen quality for culture.

Urine specimens were obtained from 115 dogs. Each specimen was divided into 2 aliquots; one aliquot was placed into a sterile container (non-preserved), and the other was preserved in a boric acid-glycerol-sodium formate at 4 C. Both aliquots were then transported to a laboratory, and specimens of each aliquot were bacteriologically cultured immediately upon arrival at the laboratory. Specimens of the preserved aliquot also were cultured after 24, 48, and 72 hours of preservation. In 47 specimens containing greater than or equal to 1 X 10(4) colony-forming units (CFU)/ml, the results of quantitative bacteriologic culturing of preserved urine at 0, 24, 48, and 72 hours were the same as the result of the immediate quantitative bacteriologic culture of non-preserved urine. In 2 of 5 specimens containing greater than or equal to 1 X 10(3) CFU/ml but less than 1 X 10(4) CFU/ml, the results of quantitative bacteriologic culture of preserved urine differed from the corresponding immediate culture of nonpreserved urine. Bacteria were not isolated from 63 specimens (less than 1 X 10(3) CFU/ml). Bacteriologic cultures of preserved urine were also negative for bacterial growth at 0, 24, 48, and 72 hours. Bacteriuria was detected by direct microscopic examination of gram-stained smears of uncentrifuged urine in 50 of 52 urine specimens from dogs with urinary tract infection. Bacteria were observed in 1 of 63 specimens that did not have bacterial growth when cultured.

Animals↗

PubMed1985

Efficacy of chlorhexidine cleansing in reducing contamination of bagged urine specimens.

To determine the effectiveness of precleansing with chlorhexidine gluconate-cetrimide in reducing the contamination rate of bagged urine specimens, 62 infants admitted to a children's hospital were randomly assigned to either receive (32 infants) or not receive (30) cleansing before bag application. Perimeatal swabs were taken before bag application and, in the treated group, after cleansing. Of the specimens from the treated group 69% were found to be contaminated, compared with 73% of those from the no-cleansing group. Chlorhexidine was ineffective in eliminating the perimeatal flora in 75% of the infants. The same organisms were present on the perimeatal swab and in the urine specimen in 95% of the infants in the treated group and 96% of those in the no-cleansing group. To estimate the contamination rate of urine specimens routinely cultured in the laboratory, 200 consecutive specimens (142 midstream and 58 bagged) were cultured. The contamination rate of the midstream urine specimens was 15%, compared with 66% for the bagged specimens. The cost of laboratory processing of contaminated bagged urine specimens at the hospital in 1983 may have been as high as $13 365. Chlorhexidine cleansing does not appear to be cost-effective. Further randomized controlled studies are needed to evaluate the effectiveness of other cleansing agents in reducing the contamination rate of bagged urine specimens.

Chlorhexidine↗

PubMed1997

Mammographically directed breast biopsies: a College of American Pathologists Q-Probes study of clinical physician expectations and of specimen handling and reporting characteristics in 434 institutions.

OBJECTIVES: To determine (1) the information that clinical physicians want in surgical pathology reports of biopsies performed for mammographic abnormalities, (2) how specimens are received and processed, and (3) what information is actually present in pathology reports. DESIGN: The standard data collection and questionnaire format of the College of American Pathologists' Q-Probes quality improvement program was used. Laboratories (1) surveyed clinical physicians working in selected specialties involved in the care of patients with breast disease concerning what information they wanted in a surgical pathology report, (2) documented laboratory specimen handling for 20 consecutive breast biopsy tissues obtained to investigate mammographic abnormalities, and (3) documented the information content of surgical pathology reports of these cases. PARTICIPANTS: Four hundred thirty-four participating laboratories surveyed 1469 clinical physicians and collected information on 7300 cases regarding specimen processing and report content. RESULTS: Clinical physicians were unanimous in the majority of items they desired for patient care. The information, however, differed depending on the diagnosis. In processing of tissues, 89% of specimens were received without fixative. In 83% of cases, radiographs were performed. Eighty-two percent of specimens were marked in some manner, but in only 45% of cases was a report of the radiographic abnormality given to the Pathology Department. The median number of blocks used to sample the lesion and the whole specimen was three and six blocks, respectively. The latter correlated with specimen size. Fifty-seven (13%) surgical pathology laboratories had radiography equipment. Radiographs of specimens and tissue blocks were made in 5% and 4% of cases, respectively. Correlation of a mammographic abnormality with a microscopic finding was documented in 62% of reports. In 92% of malignant cases the margin status was reported, and 77% of reports contained the lesion size. Eighty-three percent of reports with invasive carcinoma stated the tumor grade, and 76% stated the extent of intraductal carcinoma. The percentage of reports containing information items was significantly higher (P < .05) for institutions using checklists. SUMMARY: This multi-institutional Q-Probes study describes the current clinical expectations and laboratory practices associated with mammographically directed biopsies. Disparities between pathology reporting and clinician desires, as well as radiology and laboratory specimen handling practices, have been identified and may help focus future quality improvement efforts.

Biopsy↗

PubMedDate not supplied

The adequacy of capillary specimens for determining whole blood lead.

In response to demands for reliable alternatives to collection of venous specimens for determination of whole blood lead levels in children, the Centers for Disease Control has called for increased research into capillary methodologies. In this study, a three tiered approach was developed to assess the adequacy of capillary specimens for determining whole blood lead. Patient blood lead results from capillary and venous specimens were compared for obvious differences. Next, follow-up specimens for patients with elevated lead levels were compared with the initial results. In addition, experiments were conducted to determine whether or not handwashing eliminates gross contamination. Although the differences are not clinically important, the mean, 3.83 micrograms/dL for 5,100 venous specimens, was significantly lower (p < 0.005) then the mean of 4.6 micrograms/dL for 1,100 capillary specimens. Gross contamination was rare. Lead levels in follow-up specimens on patients whose initial screens were elevated were generally low. Handwashing greatly reduced the amount of external lead contamination. It is concluded that capillary specimens are an acceptable alternative to venous specimens for whole blood screening programs provided the patient and collector meticulously follow the prescribed collection protocol. Nevertheless, all elevated whole blood lead screening results, venous or capillary, should be confirmed with a venous collection before follow-up action is taken.

Blood Specimen Collection↗

PubMed1996

Monoclonality of B-cell lineage in primary pulmonary lymphoma demonstrated by immunoglobulin heavy chain gene sequence analysis of histologically non-definitive transbronchial biopsy specimens.

Immunoglobulin heavy chain (IgH) gene rearrangements were amplified in transbronchial lung biopsy (TBLB) specimens taken from five patients with primary lymphoma of the lung in whom the diagnosis was established by surgical specimens. By histopathological analysis of TBLB specimens, only two of the five cases were diagnosed as lymphoma, the other three cases being classified as equivocal due mainly to low levels of cellular atypia and to artefactual distortion. All five TBLB specimens, as well as the subsequent surgical specimens, showed a sharp monoclonal band of IgH gene rearrangement on electrophoresis of polymerase chain reaction (PCR) products. By contrast, three surgical biopsy specimens from cases of lymphoid interstitial pneumonia (LIP) showed smear polyclonal bands. No clonal rearrangements were detected in six non-neoplastic controls, including five cases of chronic bronchitis and one of sarcoidosis. The PCR products of three of the lymphoma cases were sequenced from both TBLB and surgical specimens. In all three cases, there was dominant expression of a particular rearrangement, assumed to be tumour-derived. In each case, the major clones derived from the TBLB and the surgical specimen were identical. In both lymphoma and LIP cases, more frequent usages of JH4 and JH6 were evident. The diagnosis of lymphoma can be confirmed on TBLB specimens by use of this technique.

Adult↗

PubMed1997

Detection of chromosomal aberrations in cytologic brush specimens from head and neck squamous cell carcinoma.

BACKGROUND: Detection of genetic changes in the mucosa of the upper aerodigestive tract may provide a target for the screening of cytologic specimens to identify premalignant transformation in this region. In this pilot study, the feasibility of the fluorescence in situ hybridization (FISH) technique to detect genetically aberrant cells in brush specimens was evaluated. METHODS: Brush specimens taken from the tumors of 20 patients with head and neck squamous cell carcinoma (HNSCC) and from the normal mucosa of 8 control patients were analyzed by FISH using DNA probes for the chromosomes 1 and 7. The FISH results were compared with DNA flow cytometry and FISH results of the solid tumor specimens. RESULTS: The results of this study showed that 15 of the 20 tumor brush specimens contained numeric chromosomal aberrations in at least 5% of the cells collected. Chromosomal aberrations were detected in all brush specimens taken from tumors that were DNA aneuploid and showed aneusomy. The presence of these aberrations correlated well with the classification "suspicious for malignancy," which was based on Papanicolaou stained slides of the same specimens. In the control group the percentage of chromosomally aberrant cells did not exceed 2%; in addition, no suspiciously malignant cells were observed in this group. CONCLUSIONS: This study reveals that the FISH technique can be applied diagnostically to brush specimens of HNSCC. The presence of chromosomal aberrations in > 5% of the cells in these specimens can be considered as a marker for malignancy.

Adult↗