Quality Of Smears Research Articles

An Evaluation of the National Malaria Surveillance System of Bhutan, 2006-2012 as It Approaches the Goal of Malaria Elimination.

Bhutan is progressing toward malaria elimination. The purpose of this evaluation was to assess the ability of the surveillance system from 2006 to 2012 to meet the objectives of the Bhutan Vector-borne Disease Control Program (VDCP) and to highlight priorities requiring attention as the nation transitions to elimination. The evaluation was conducted using the Center for Disease Control guidelines for evaluating public health surveillance systems. Data sources included a search of publically available literature, VDCP program data, and interviews with malaria surveillance personnel. Blood slide quality assurance and control through formal assessment of slide preparation and measures of between-reader correlation were performed. Total malaria cases declined from 2006 to 2012. The average slide positivity rate decreased from 3.4% in 2006 to 0.2% in 2012. The proportion of non-residents in all cases increased to its highest value of 22.6% in 2012, and significant clustering in the border regions of India was noted, with Sarpang accounting for more cases than any other district from 2009 onward. Case detection was almost exclusively passive, but flexibility and sensitivity was demonstrated by the later addition of active case detection and specification of imported and locally acquired cases. Spatial data were limited to the village level, not allowing identification of transmission hotspots. For blood smears, statistical measures of between-reader agreement and predictive value were not computed. Blood smear quality was suboptimal by at least one criterion in over half of evaluated smears. Timeliness in reporting of cases was on a weekly to monthly basis, and did not meet the WHO goal of immediate notification. As of 2012, the national malaria surveillance system demonstrated flexibility, representativeness, simplicity, and stability. The full potential for data analysis was not yet realized. Attaining the goal of malaria elimination will require system function enhancement through increased and more accurate case detection and rapid investigation, improved health worker training and accountability, focally targeted response measures, and, in particular, the challenge of finding re-introductions of infections from India. Many such measure have been undertaken or planned as part of the next phase of the Bhutan's National Strategic Plan.

Importance of quality control of baciloscopy in laboratories that perform diagnosis of tuberculosis

Baciloscopy is the primary tool for pulmonary tuberculosis diagnosis, being this technique the most used internationally in the search for infectious cases. Quality control is the process of the rechecking smears by a highly qualified observer. To evaluate and highlight the importance of quality control of smear microscopy in the Provincial Laboratories diagnosticians of Tuberculosis in Cuba. This study was conducted at the National Reference Laboratory and Research in Tuberculosis, Leprosy and Mycobacteria in the Institute of Tropical Medicine "Pedro Kouri", Havana, Cuba, Were evaluated 2676 smears received from January 2013 to December 2014, from Provincial Centers of Hygiene, Epidemiology and Microbiology of Cuba, including the special municipality Isla de la Juventud. 2,664 (99.5%) were concordant smears, the correlation obtained for the positive smears were 96.5% and 99.8% for negative. Were identified12 reading errors: 7 (3.5%) false positive and 5 (0.2%) false negatives. Slides were classified with adequate quality of smears in 2039 (76.2%), showed difficulties in realizing the extension in 1464 (54.7%) and staining were adequate in 2343 (87.6%). The kappa index was 0.9674. Although there was good agreement between observations it is recommended to improve the quality of extended, maintain staff training program that performs this activity, like regular supervision by specialists, to further improve the quality of diagnosis.

Decentralization of Acid Fast Bacilli(AFB) External Quality Assurance Using Blind Rechecking for Sputum Smear Microscopy in Ethiopia

IntroductionEthiopia achieved a rapid expansion of TB microscopic centers for acid fast bacilli (AFB). However, external quality assurance (EQA) services were, until recently, limited to few regional and sub-regional laboratories. In this paper, we describe the decentralization experience and the result of EQA using random blinded rechecking.Materials and MethodsThe routine EQA quarterly report was compiled and analyzed. A positive result by the microscopic center while the EQA center reported negative result is categorized as false positive (FP). A negative result by the microscopic center while the EQA center reported positive is considered false negative (FN). The reading of EQA centers was considered a gold standard to compute the sensitivity, specificity, positive predictive (PPV) and negative predictive values (NPV) of the readings of microscopic centers.ResultsWe decentralized sputum smear AFB EQA from 4 Regional Laboratories (RRLs) to 82 EQA centers and enrolled 956 health facilities in EQA schemes. Enrollment of HFs in EQA was gradual because it required training and mentoring laboratory professionals, institutionalizing internal QA measures, equipping all HFs to perform diagnosis, and establishing more EQA centers. From 2012 to 2014 (Phase I), the FP rate declined from 0.6% to 0.2% and FN fell from as high as 7.6% to 1.6% in supported health facilities (HFs). In HFs that joined in Phase II, FN rates ranged from 5.6 to 7.3%. The proportion of HFs without errors has increased from 77.9% to 90.5% in Phase I HFs and from 82.9% to 86.9% in Phase II HFs. Overall sensitivity and specificity were 95.0% and 99.7%, respectively. PPV and NPV were 93.3% and 99.7%, respectively.ConclusionDecentralizing blinded rechecking of sputum smear microscopy is feasible in low-income settings. While a comprehensive laboratory improvement strategy enhanced the quality of microscopy, laboratory professionals’ capacity in slide reading and smear quality requires continued support.

FNA smears as a potential source of DNA for targeted next-generation sequencing of lung adenocarcinomas.

Diff-Quik-stained fine-needle aspiration (FNA) smears and touch preparations from biopsies represent alternative specimens for molecular testing when cell block or biopsy material is insufficient. This study describes the use of these samples for targeted next-generation sequencing (NGS) of primary and metastatic lung adenocarcinoma and reports the DNA quality and success rates of FNA smears versus other specimens from 1 year of clinical use. A validation set of 10 slides from 9 patients with prior clinical epidermal growth factor receptor (EGFR) Sanger sequencing and KRAS pyrosequencing (5 KRAS-positive/EGFR-negative and 4 KRAS-negative/EGFR-negative) underwent DNA extraction, quality assessment, and targeted NGS. Subsequently, lung adenocarcinoma specimens submitted for NGS solid tumor mutation panel testing in 1 calendar year (60 biopsies, 57 resections, 33 FNA cell blocks, 12 FNA smears, and 10 body fluid cell blocks) were reviewed for specimen adequacy, sequencing success, and DNA quality. All 10 validation samples met the DNA quality threshold (delta Ct threshold < 8; range, -2.2 to 4.9) and yielded 0.5 to 22 μg of DNA. The KRAS and EGFR mutation status from FNA smears according to NGS was concordant with previous clinical testing for all 10 samples. In the 1-year review, FNA smears were 100% successful, and this suggested a performance equivalent to or better than the performance of established specimen types, including FNA cell blocks. DNA quality according to ΔCt was significantly better with FNA smears versus biopsies, resections, and FNA cell blocks. FNA smears of lung adenocarcinomas are high-quality alternative specimens for a targeted NGS panel with a high success rate in clinical practice. Cancer Cytopathol 2016;124:406-14. © 2016 American Cancer Society.

Quality analysis of malaria blood smears of fever patients in Changzhou City in 2014

To understand the quality of blood smears for Plasmodium detection in Changzhou City, so as to provide the technical support for malaria surveillance in the post-elimination period. The blood smears of fever patients from each cities (districts) in Changzhou City in 2014, including more than 3% of the negatives and all the positives, were selected. The quality of the blood smear making, dyeing, cleanliness and results were reviewed by the experts from the municipal malaria microscopy station, and the data were analyzed statistically. Totally 996 negative blood smears were reviewed in 2014, with a review rate of 4.52%. The qualified rates of blood smear making, dyeing and cleanliness were 92.87%, 93.27% and 94.48%, respectively. A total of 34 positive blood smears were reviewed, and there were no false detections and leak detections. The qualified rates of blood smear making and dyeing were more than 90% in all the seven cities (districts). As for qualified rate of cleanliness, except the rate in Qishuyan District (81.36%), those in the other six cities (districts) were all above 90%. The qualified rates of blood smear making, dyeing and cleanliness were all more than 90% in first-class, second-class and third-class hospitals. The major quality defects of the blood smears were sediment, abnormal making and thick membrane shedding, and the proportions were 25.91%, 21.76% and 19.17%, respectively. The quality of blood smears for Plasmodium detection in Changzhou City is satisfying. In the future, the training and supervision still should be strengthened to guarantee the capacity of microscopic examinations for Plasmodium detection in the surveillance of post-elimination period.

Hallazgos del lavado broncoalveolar procesado con técnica de cytospin y citología convencional

The cytological interpretation of bronchoalveolar lavage (BAL) specimen is currently performed using conventional cytology and cytospin preparations. The latter features concentrated preparations in a 6-mm area for sample visualisation and reduced reading time, with no deterioration in quality. ObjectiveTo determine the differences in reading time and quality, between the conventional technique and the cytospin technique. Study designA cross-sectional study was conducted on BAL specimens that were processed using the conventional technique and the cytospin technique. Reading time for diagnosis between the 2 techniques was compared. Quality was assessed taking into account, cell preservation and inflammatory cells in smears. The difference in time was calculated using the Wilcoxon test. ResultsEighty (80) BAL slides were examined, 40 by the conventional technique and 40 by the cytospin technique. A differential count was not performed in 6 cases, in 4 due to excessive number of squamous cells and in 2 cases, processed by the conventional technique, due to exhibiting less than 300 cells. A marked degeneration of epithelial cells was observed using the conventional technique, showing a significant difference in reading time (P<.001). ConclusionsA significant difference was found between reading times of the 2 techniques, which was longer when using the conventional technique. Better quality smears were observed when using the cytospin technique.

PAPILLOMA VIRAL INFECTION IN PREGNANT WOMEN: WAYS TO SOLVE THE PROBLEM

Peculiarities of the course of the papilloma viral infection in pregnancy are studied insufficiently. There is no generally accepted practice of patient maintenance. The aim of the study is to study possibilities of pharmaceutical correction of the papilloma viral infection (PVI) affected pregnant women. Various types of human papilloma virus are identified in 137 patients by polymerase chain reaction method. The immune system correction and therapy of sexually transmitted diseases was administered to 33 women. 31 women didn’t receive the immune system correction of therapy of sexually transmitted diseases. Spiramycyin, natamycyin, bifidumbacterin, tergynan, inferferon alpha2b + taurin were used for therapy. In addition to clinical and bacteriological study methods extended uterine colposcopy and cytologic methods of cervical smear were used. The obtained results allowed drawing a conclusion that the sexually transmitted diseases drug therapy and the immune system correction, improving the colposcopic pattern and quality of cytologic smears optimizing the course of pregnancy; the sexually transmitted disease therapy is less effective than simultaneous sexually transmitted disease therapy and immune system correction.

Choropleth Mapping of Cervical Cancer Screening in South Africa Using Healthcare Facility-level Data from the National Laboratory Network.

BackgroundIn South Africa, cervical cancer remains among the most common cancers and a leading cause of cancer death. Co-infection with HIV increases the risk of developing cervical pre-cancer and cancer. We analysed National Health Laboratory Service cervical cytology data to investigate geographic variations of Pap smear coverage, quality, and high grade lesions.MethodsFacility-level data were extracted from the NHLS for April 2013–March 2014. We present results and choropleth maps detailing coverage, adequacy and high-grade Pap smear cytology abnormalities defined as Pap smears suspicious for invasive carcinoma, high-grade squamous intraepithelial lesions (HSIL) or atypical squamous cells: cannot exclude HSIL (ASC-H).Results4,562 facilities submitted 791,067 cytology slides. The interquartile range (IQR) for Pap smear coverage among HIV-infected women was 26–41%; similar to coverage in women aged 30 and older (IQR: 26–42%). 6/52 districts had adequacy rates above the national standard (70%) and 2/52 districts had adequacy rates below 35%. We observed marked variation in Pap smear abnormalities across the country, with the proportion of high-grade cytology abnormalities ≥0.3% in 17/52 districts.ConclusionUsing district-level choropleth maps, we are able to display variations in Pap smear coverage, quality, and results across South Africa. This approach may be used to improve resource allocation, achieving better equity in cervical cancer prevention.

Tuberculosis Laboratory Diagnosis Quality Assurance among Public Health Facilities in West Amhara Region, Ethiopia.

IntroductionReliable smear microscopy is an important component of Directly Observed Treatment Scheme (DOTS) strategy for TB control program in countries with limited resources. Despite external quality assessment is established in Ethiopia, there is lower TB detection rate (48%) in Amhara region compared to the World Health Organization (WHO) estimate (70%). This highlights the quality of smear microscopy needs to be evaluated. Therefore, the aim of this study was to assess the quality of sputum smear microscopy performance among health center laboratories in West Amhara region, Ethiopia.Materials and MethodsA cross sectional study was conducted from July 08, 2013 to July 07, 2014. Data were collected from 201 public health center laboratories using a structured questionnaire. Slides were collected based on Lot Quality Assurance Sampling (LQAS) method and rechecked blindly by trained laboratory technologists. The data were entered into EPI info V.7 and smear quality indicators and AFB results were analyzed by SPSS version 20.ResultsAmong 201 laboratories enrolled in this study, 47 (23.4%) laboratories had major errors. Forty one (20.4%) laboratories had a total of 67 false negative and 29 (14.4%) laboratories had a total of 68 false positive results. Specimen quality, smear thickness and evenness were found poor in 134 (66.7%), 133 (66.2%) and 126 (62.7%) laboratories, respectively. Unavailability of microscope lens cleaning solution (AOR: 2.90; 95% CI: 1.25–6.75; P: 0.013) and dirty smears (AOR: 2.65; 95% CI: 1.14–6.18; P: 0.024) were correlated with false negative results whereas no previous EQA participation (AOR: 3.43; 95% CI: 1. 39-8.45; P: 0.007) was associated with false positive results.ConclusionThe performance of health facilities for sputum smear microscopy was relatively poor in West Amhara region. Hence, strengthening the EQA program and technical support on sputum smear microscopy are recommended to ensure quality tuberculosis diagnostic service.

A novel cytologic sampling technique to diagnose subclinical endometritis and comparison of staining methods for endometrial cytology samples in dairy cows

The present article describes a study of the diagnosis of subclinical endometritis in dairy cows having two principal aims: first, to validate a novel technique for taking endometrial cytology samples to diagnose subclinical endometritis in dairy cows. Second, to compare the percentage of polymorphonuclear cells (PMNs) in cytology samples stained with Diff-Quik versus a specific staining method for PMNs, naphthol-AS-D-chloroacetate-esterase (CIAE). In the first experiment, Holstein-Friesian cows (n = 204) were used to take two cytology samples at the same time using the conventional cytobrush (CB) and the new cytotape (CT). Both devices were assembled within the same catheter allowing sampling at the same time, and approximately at the same location. Cytotape consisted of a 1.5-cm piece of paper tape rolled on the top of an insemination catheter covered with a double guard sheet. Parameters used to evaluate both methods were: PMNs percentage, total cellularity, quality of the smears, and red blood cell contamination. The concordance correlation coefficient analysis was used to assess agreement between continuous and Pearson chi-square tests for categorical variables. Agreement between the percentage of PMNs in both methods was good ρ = 0.84 (0.79, 0.87) with a minor standard error of 2%. Both methods yielded similar total cellularity (P = 0.62). Cytotape yielded better quality smears with more intact cells (P < 0.01) while samples that were taken by CB were more likely to be bloody (P < 0.01). Hence, CT and CB methods yielded smears with a similar PMNs percentage and a total number of cells, but CT provided smears with higher quality and significantly less blood contamination. For the second experiment, 114 duplicate cytology slides were stained using both Diff-Quik and CIAE. Agreement between PMNs percentage in both staining techniques was good ρc = 0.84 (0.78, 0.89) with a standard error of only 2%. Hence, Diff-Quik was confirmed as an easy, fast, and high-quality staining technique, which can be routinely used to stain endometrial cytology samples satisfactorily.

Comparison of specimen adequacy and smear quality in oral smears prepared by manual liquid-based cytology and conventional methods.

Background:Liquid-based cytology (LBC), recommended in the mass screening of potentially malignant cervical and oral lesions, suffers from high cost owing to the use of expensive automated devices and materials. Considering the need for cost-effective LBC techniques, we evaluated the efficacy of an inexpensive manual LBC (MLBC) technique against conventional cytological technique in terms of specimen adequacy and smear quality of oral smears.Materials and Methods:Cytological samples were collected from 21 patients using a cytobrush device. After preparation of a conventional smear, the brush containing the remaining sample was immersed in the preservative vial. The preserved material was processed by an MLBC technique and subsequently, direct smears were made from the prepared cell button. Both conventional and MLBC smears were stained by routine Papanicolaou technique and evaluated by an independent observer for the thickness of the smear, cellular distribution, resolution/clarity of cells, cellular staining characteristics and the presence of unsatisfactory background/artifacts. Each parameter was graded as satisfactory; or satisfactory, but limited; or unsatisfactory. Chi-square test was used to compare the values obtained (significance set at P ≤ 0.05).Results:MLBC technique produced a significant number of satisfactory smears with regard to cell distribution, clarity/resolution, staining characteristics and background/artifacts compared to conventional methods.Conclusions:MLBC is a cost-effective cytological technique that may produce oral smears with excellent cytomorphology and longer storage life.

Fine Needle Aspiration Cytology versus Fine Needle Capillary Sampling in Cytological Diagnosis of Thyroid Lesions.

Fine needle aspiration cytology (FNAC) is an established out- patient procedure used in primary diagnosis of palpable thyroid lesions. A modified technique fine needle capillary sampling (FNCS) obviates the need of suction, is less painful, patient friendly and reported to overcome the problem of inadequate and bloody specimens. The aim of our study was to compare the efficacy and quality of FNCS with that of conventional FNAC in the lesions of thyroid. One hundred patients, presenting between January 2011 to December 2012 at Cytopathology Department of M M Institute of Medical Sciences and Research, Mullana, with diffuse and nodular thyroid lesions were enrolled with both the techniques being executed on the patients, beginning with FNA followed by FNCS. The smears were scored using five objective parameters i.e. background blood, cellular material, cellular degeneration, cellular trauma, and retention of appropriate architecture, in a single blind setting by a cyto-pathologist. The results were analyzed using Student's test for paired data and chi- square analysis. A highly significant differences (P<0.001) in favor of FNCS was observed for the background blood, cellular material and retention of architecture while total score favored FNA for cellular degeneration and degree of cellular trauma. Total scores and average score per case for FNCS was significantly better (P<0.001) than FNA. FNCS technique yielded more diagnostically superior and lesser number of unsatisfactory smears whereas greater number of diagnostically adequate samples was obtained by FNA technique. FNCS offers more number of diagnostically better quality smears. Both techniques could be supplementary on many occasions and substitutive on a few. Combination of the two techniques could offer better diagnostic accuracy.

Spontaneous and induced sputum values in the diagnosis of pulmonary tuberculosis among patients with suspected pulmonary tuberculosis

Objectives: The quality of sputum samples is important for the diagnosis of pulmonary tuberculosis (TB). Unfortunately, it is not always possible to get a proper sputum sample. Now, sputum induction (SI) has been found to be useful for the diagnosis of many lung diseases. Therefore, this study compares the role of spontaneous sputum and induced sputum in the diagnosis of pulmonary TB among suspected pulmonary TB patients. Materials and Methods: A total of 27 patients admitted with suspected pulmonary TB were studied. Three spontaneous sputum samples were taken from these patients and in case of negative sputum smear microscopy, the SI procedure was used to take another sputum sample. Standard diagnosis included both positive sputum smear and clinical decisions. The sensitivity, specificity, and positive and negative predictive values of sputum smears obtained by two methods of spontaneous sputum and induced sputum were calculated and compared with the standard diagnosis. Results: The result of the induced sputum acid-fast bacillus (AFB) smear microscopy was negative in 16 (59.3%) patients and positive in 11 (40.7%) patients. Six (50%) of those who were negative in spontaneous sputum became positive after SI. The mean white blood cell (WBC) count in normal sputum and induced sputum was 3.8 (±3.4) and 8.8 (±2.9), respectively (P < 0.001). The sensitivity of sputum smear positive was calculated to be 67.7% for SI method and 29.4% for spontaneous sputum method. Conclusion: The diagnostic value of induced sputum in the diagnosis of TB is more than the spontaneous sputum method. In addition, the quality of sputum smear produced through induction is more than spontaneous sputum and it does not create any special complication.

Microscopy for Acid Fast Bacilli: A Useful but Neglected Tool in Routine Laboratory Diagnosis of Buruli Ulcer

Background: Laboratory diagnosis of Buruli ulcer disease has become vital with the introduction of antibiotic treatment. Polymerase chain reaction (PCR) for the IS2404 repeat sequence of Mycobacterium ulcerans is the gold standard for laboratory diagnosis. This is expensive and only carried out in reference laboratories in endemic countries in Africa. In order to improve the efficiency of diagnosis at the point of care and reduce the total cost of patient management, we decided to evaluate Ziehl-Neelsen (ZN) staining for acid-fast bacilli (AFB) as an inexpensive diagnostic tool. Methods: Two smears directly prepared at the point of care were examined under oil-immersion microscopy after ZN staining for AFB and compared the results to PCR samples from the same patients. Results: Good quality smears were obtained from all subjects and our results showed that when a second smear was added the sensitivity of microscopy for AFB was increased from 52 to 55% for FNA samples and from 51 to 57 % for swabs. Conclusion: If PCR were to be omitted in all patients with suspected Buruli ulcer disease when AFB were detected it would result in a considerable saving.

Impact of a training course on the quality of malaria diagnosis by microscopy in Angola.

BackgroundIn Angola, malaria is an endemic disease having a major impact on the economy. The WHO recommends testing for all suspected malaria cases, to avoid the presumptive treatment of this disease. In malaria endemic regions laboratory technicians must be very comfortable with microscopy, the golden standard for malaria diagnosis, to avoid the incorrect diagnosis. The improper use of medication promotes drug resistance and undesirable side effects. The present study aims to assess the impact of a three-day refresher course on the knowledge of technicians, quality of blood smears preparation and accuracy of microscopy malaria diagnosis, using qPCR as reference method.MethodsThis study was implemented in laboratories from three hospitals in different provinces of Angola: Bengo, Benguela and Luanda. In each laboratory samples were collected before and after the training course (slide with thin and thick blood smears, a dried blood spot and a form). The impact of the intervention was evaluated through a written test, the quality of slide preparation and the performance of microscopy.ResultsIt was found a significant increase on the written test median score, from 52.5% to 65.0%. A total of 973 slides were analysed to evaluate the quality of thick and thin blood smears. Considering all laboratories there was a significant increase in quality of thick and thin blood smears. To determine the performance of microscopy using qPCR as the reference method we used 1,028 samples. Benguela presented the highest values for specificity, 92.9% and 98.8% pre and post-course, respectively and for sensitivity the best pre-course was Benguela (75.9%) and post-course Luanda (75.0%). However, no significant increase in sensitivity and specificity after the training course was registered in any laboratory analysed.DiscussionThe findings of this study support the need of continuous refresher training for microscopists and other laboratory staff. The laboratories should have a quality control programme to supervise the diagnosis and also to assess the periodicity of new training. However, other variables needed to be considered to have a correct malaria diagnosis, such as adequate equipment and reagents for staining and visualization, good working conditions, motivated and qualified personnel.

The quality of cervical smears from outreach screening services in remote areas in eastern Taiwan

ObjectivesIssues such as specimen adequacy and diagnostic yield have rarely been studied in rural regions. This study investigated specimen adequacy and the diagnostic yield of cervical smears from outreach screening services in remote regions in eastern Taiwan. Materials and methodsThere were 5670 smears from outreach screening services; 649 smears were taken in mobile screening van units or in temporary walk-in clinics. The quality, compromising factors, and positive rates of smears from mobile screening van units and temporary walk-in clinics were compared with those obtained in physician offices. ResultsThe quality of smears from physician offices was better than the quality from mobile van screening units and temporary walk-in clinics; the satisfactory rates were 72%, 54%, and 53%, respectively (χ2 = 109.9910, p < 0.0001; df = 4). The most important limiting factor, absence of endocervical component (ECC), was reported in 56%, 57%, and 73% of suboptimal smears at physician offices, mobile van screening units, and temporary walk-in clinics, respectively (χ2 = 25.2780, p < 0.001; df = 2). Poor fixation compromised 18%, 18%, and 14% of smears taken at physician offices, mobile van screening units, and temporary walk-in clinics, respectively. For smears taken in physician offices, the odds ratio for positivity of suboptimal smears with ECC was nine times the odds ratio of suboptimal smears with no ECC. ConclusionThe issue of quality of specimens from outreach services in rural areas needs to be emphasized to improve the sensitivity of the test. In future studies, the difficulties that health workers encounter when taking smears in outreach services and the underlying sociocultural and demographic barriers need to be elucidated.

Pap Smear Advocacy in LMICs: Targeted Education to Providers Drastically Improves Pap Smear Quality and Acceptability of Cytology Based Screening in Rural South India in a Very Short Time

Pap Smear Advocacy in LMICs: Targeted Education to Providers Drastically Improves Pap Smear Quality and Acceptability of Cytology Based Screening in Rural South India in a Very Short Time

Making mock-FNA smears from fresh surgical pathology specimens to improve smear preparation technique and to create cytohistological correlation series.

BackgroundFine needle aspiration (FNA) cytology is a well-established diagnostic method based on the microscopic interpretation of often scant cytological material; therefore, experience, good technique and smear quality are equally important in obtaining satisfactory results.Aims of StudyWe studied the use of fresh surgical pathology specimens for making so-called mock-FNA smears with the potential of cytohistological correlation. Additionally, we studied how this process aids the improvement of preparation technique and smear quality.MethodsCytological aspirates from 32 fresh biopsy specimens from various sites: lung (20), lymph nodes (6), and breast (6) were obtained, all with a clinical diagnosis of tumor. Aspiration was performed from grossly palpable tumors. 25G needle and Cameco-type syringe holder was used with minimal or no suction.ResultsUnfixed surgical specimens provided sufficient cytological material that resulted in good quality smears. After standard processing of specimens into microscopic sections from paraffin embedded tissues, cytohistological case-series were created. No significant alteration was reported in tissue architecture on hematoxylin-eosin stained sections after the aspiration procedure. A gradual, but steady improvement was observed in smear quality just after a few preparations.Discussions and ConclusionsOur study proved that surgical specimens may be used as a source of cytological material to create cytohistological correlation studies and also to improve FNA cytology skills. The use of very fine gauge needle (25G, 0,6 mm diameter) during the sampling process does not alter tissue architecture therefore the final histopathological diagnosis is not compromised. We conclude that by using fresh surgical specimens useful cytohistological collections can be created both as a teaching resource and as improving experience.

Effects of modifying the World Health Organization standard operating procedures for malaria microscopy to improve surveillance in resource poor settings.

BackgroundIndividuals with fever are screened for malaria in specially-established malaria diagnostic laboratories set up in rural hospitals in the Northern and Eastern Provinces of Sri Lanka. Large numbers of blood smears negative for malaria parasites are being screened daily. Good quality smears are essential to maintain a high diagnostic competency among the technical staff. The modifications made to the World Health Organization (WHO) standard operating procedures to improve the quality of smears have been studied.MethodsA blinded, controlled, interventional study was conducted in 22 intervention and 21 control malaria diagnostic laboratories. Changes were made to the WHO standard operating procedure protocols to prepare, stain and examine blood smears for malaria parasite detection which were implemented in intervention laboratories. These included wipe-cleaning slides, preparing both thick and thin smears on the same slide, reversing the order of collecting blood for thick and thin smears, dry fixing thick smear for 20–25 minutes under table lamp, polishing the edge of spreader slide with sand paper and fixing the thin smear with methanol if not stained within four hours. Parameters with respect to quality of the smear as per WHO criteria were studied using randomly selected slides, and time taken for the report to be issued was recorded in both groups before and after the intervention.ResultsThere were no significant differences observed in the parameters studied at baseline between the two groups or pre and post intervention in the control group. In the intervention group streak formation in thin smears was reduced from 29.4% to 5.0%. The average fixing time of thick smears was reduced from 2.4 hours to 20 minutes. Inappropriate thickness of thick smears reduced from 18.3% to 1.5%. Overall quality of thick smears and thin smears increased from 76.1% to 98.0% and 81.7% to 87.0%, respectively. The quality of slides bearing both thick and thin smears increased from 60.0% to 87.0%.ConclusionsNew protocols with amendments to the WHO standard technical procedures ensure that good quality blood smears are prepared rapidly to diagnose malaria and the time required to issue the reports was reduced.

Automated Analysis of Bone Marrow Aspirates from Dogs with Haematological Disorders

Automated analysis of bone marrow (BM) aspirates is a useful 'pre-microscopical' screen to identify hypocellular samples and those with potentially abnormal cells. In order to determine whether automated analysis could also be used to identify haemopoietic abnormalities, EDTA-anticoagulated BM aspirates from 43 dogs were analysed using the Advia 2120 instrument. Corresponding Wright-stained BM smears were evaluated microscopically to determine smear quality, cell composition and 500-cell differential counts, and correlation to automated analysis parameters was computed. Leucocyte cytograms generated by the automated analyzer were scrutinized and compared with those of 'normal' BM. Twenty-three neoplastic and 20 non-neoplastic samples were analysed, including samples from 10 cases of acute myeloid leukaemia, four cases of acute lymphocytic leukaemia, four cases of chronic lymphocytic leukaemia, one case of chronic neutrophilic leukaemia, three cases of multiple myeloma, one case of myelodysplastic syndrome, five cases of non-regenerative immune-mediated haemolytic anaemia, one case of immune-mediated neutropenia, three cases of immune-mediated thrombocytopenia, six cases of inflammatory disease, three samples with myelotoxicity and two samples analysed for staging of neoplasia. Automated white blood cell (WBC) counts correlated significantly with smear cellularity, particle cellularity and particle number. There was a significant difference in WBC counts of samples with insufficient versus sufficient particles. Significant correlations between Advia percent neutrophils and microscopical determination of marrow segmented neutrophils/neutrophilic granulocyte reserve, Advia percent lymphocytes and microscopical determination of lymphocytes/rubricytes, Advia percent large unstained cells and microscopical determination of myeloblasts/promyelocytes and between Advia percent eosinophils and manual determination of eosinophils were identified. This suggested that Advia WBC counts may be used to approximate BM sample quality and that Advia differential counts may predict marrow granulocyte reserve and lymphocyte/rubricyte stores. Distinct and consistent alterations in cytogram patterns were observed in cases of acute leukaemia, but were less obvious in chronic leukaemia. Complete automated BM analysis was performed in approximately 2 min, while staining and coverslipping of BM slides required approximately 30 min. Hence, although automated analysis should not supplant microscopical evaluation of BM, it can provide useful ancillary information in a short time and flag potentially inadequate or abnormal samples.