Evaluation of urine specimen integrity in a public health STD screening program

Detection of Chlamydia trachomatis and/or Neisseria gonorrhoeae infection in urine using molecular amplification assays has permitted institutions with limited medical facilities to offer testing for these sexually transmitted diseases (STDs). The Nebraska Public Health Laboratory (NPHL) investigated the validity of urine samples submitted for C trachomatis and/or N gonorrhoeae amplification after receiving a substantial number of clear specimens. Approximately 75% of all urine specimens submitted for STD testing to the NPHL were from correctional facilities. The falsification of urine specimens submitted for microbiology studies is not evaluated routinely, and this problem was previously undocumented. By using the criteria for specific gravity of 1.001 or less and a creatinine concentration of less than 5 mg/dL (442 mumol/L), approximately 8% of all specimens submitted during the study interval were determined to be inconsistent with urine. The microbiology laboratory should be aware of the possibility for specimen manipulation to identify facilities submitting falsified specimens, to initiate appropriate intervention, and to minimize false-negative reporting.     

The validity of random urine specimen albumin measurement as a screening test for diabetic nephropathy

To assess the validity of urine albumin concentration (UAC) and the urine albumin:creatine ratio (UACR) in a random urine specimen (RUS) for screening diabetic nephropathy in Korea, a total of 105 ambulatory diabetes mellitus patients (male:female, 52:53), ages 40-75 years (median 59 years) collected 105 RUSs after completing a timed 24 hour urine collection. Albumin was measured by immunonephelometry. According to the timed urinary albumin excretion rate (UAER) measured in the 24 hour collection (criterion standard), samples were classified normoalbuminuric (UAER < 20 micrograms/min; n = 50), microalbuminuric (UAER 20-200 micrograms/min; n = 30), and macroalbuminuric (UAER > 200 micrograms/min; n = 25). The receiver operating characteristics (ROC) curve of UAC and UACR in a RUS for screening of microalbuminuria (normo- and microalbuminuric samples; n = 80) and macroalbuminuria (micro- and macroalbuminuric samples; n = 55) were plotted. Pearson's coefficients of correlation of 24 hour UAER vs. UAC and UACR were 0.81 and 0.75, respectively (P < 0.001). The point of intersection with a 100%-to-100% diagonal for microalbuminuria were as follows: 31.0 mg/l for UAC and 32.5 mg/g for UACR; for macroalbuminuria 181 mg/l for UAC and 287.3 mg/g for UACR. The sensitivity and specificity of the cut-off points for microalbuminuria were 77% and 82% for UAC and 77% and 92% for UACR. The sensitivity and specificity of the cut-off points for macroalbuminuria were 84% and 90% for UAC and 88% and 90% for UACR. In present study, no difference was observed when comparing the performance of UAC and UACR based on a statistical comparison by McNemar test. The repeated measurements of UAC and UACR in the same individual were statistically similar and were correlated with each other. Based on these results, albumin measurements (UAC and UACR) in a RUS were considered as a valid test for screening diabetic nephropathy.     

Evaluation of a finger-stick specimen collection method for seroepidemiology of antibody to hepatitis A virus

A finger-stick swab method of collecting blood specimens was shown to compare favorably with the conventional venipuncture method in serological determinations of antibody to hepatitis A virus by radioimmunoassay.     

Evaluation of number of shell vial cell cultures per clinical specimen for rapid diagnosis of cytomegalovirus infection.

Specimens submitted for the diagnosis of cytomegalovirus (CMV) infection were inoculated into three (blood) or two (urine, tissue, bronchoalveolar lavage [BAL]) shell vials seeded with MRC-5 cells for the diagnosis of CMV infection. We evaluated the detection of 993 specimens that were positive for CMV according to the number of shell vial cell cultures inoculated per specimen. For blood cultures, and considering one CMV-positive shell vial as 100%, inoculation of three shell vials versus one increased the detection rate of the virus by 51%. Inoculation of three shell vials compared with two yielded a 20% increase in the detection rate of CMV. For urine, tissue, and BAL specimens, inoculation of two shell vials compared with one resulted in increases of 7, 10, and 5%, respectively. For maximum detection of CMV in shell vial cell cultures, at least three vials should be inoculated with blood specimens, and two vials should be used for urine, tissue, and BAL samples.     

High-grade prostatic sarcoma seen in a catheterized urine specimen: case report and differential diagnosis

Urine cytology has been effectively used in the diagnosis and management of epithelial bladder tumors, particularly high-grade urothelial carcinoma. Indeed it is the gold standard for bladder cancer screening. Although urothelial carcinoma is the most frequently identified bladder tumor by urine cytology, metastatic carcinomas from the kidney, colon, and a variety of other regional organs have been detected. Stromal lesions such as inflammatory myofibroblastic tumor, gastrointestinal stromal tumor, and leiomyosarcoma are other much rarer entities occurring within the bladder. Few to no case reports exist documenting their identification within urine cytology specimens. Herein we report the detection of a high-grade prostatic sarcoma within a catheterized urine specimen of a young male having a diffusely enlarged prostate. The specimen consisted of numerous fragments of relatively uniform spindle cells having ovoid nuclei with rounded ends and finely dispersed chromatin. Cytoplasmic borders were indistinct. No mitoses or significant atypia was present. The background consisted of numerous red blood cells, cellular debris, and a few clusters of unremarkable urothelial cells. Followup surgical biopsy of the patient's prostate revealed a high-grade spindle-cell sarcoma. Further immunohistochemical and molecular delineation of the tumor was not informative for a more definitive diagnosis. Although rare, sarcomas and other mesenchymal tumors involving the bladder are unique entities with a broad differential diagnosis.     

Serum is the preferred clinical specimen for diagnosis of human brucellosis by PCR

Human brucellosis poses a significant public health problem in many developing countries and requires fast and accurate diagnosis. A PCR assay amplifying part of the 31-kDa Brucella abortus antigenic protein gene sequence was developed and applied to whole-blood and serum samples from 31 brucellosis patients and 45 healthy individuals. All patients except one had detectable Brucella DNA in either whole blood or serum (combined sensitivity, 97%), but the assay sensitivity was higher with serum samples (94%) than with whole-blood samples (61%). The assay specificity was excellent (100%). A confirmatory PCR assay targeting another Brucella gene region (omp-2) was also developed but lacked sensitivity. Serum is the optimal specimen for the diagnosis of brucellosis by PCR, a choice that leads to assay simplification and shortens turnaround time.     

Saliva sample as a non-invasive specimen for the diagnosis of coronavirus disease 2019: a cross-sectional study

ObjectivesAmid the increasing number of pandemic coronavirus disease 2019 (COVID-19) cases, there is a need for a quick and easy method to obtain a non-invasive sample for the detection of this novel coronavirus (severe acute respiratory syndrome coronavirus 2; SARS-CoV-2). We aimed to investigate the potential use of saliva samples as a non-invasive tool for the diagnosis of COVID-19.MethodsFrom 27 March to 4 April 2020, we prospectively collected saliva samples and a standard nasopharyngeal and throat swab in persons seeking care at an acute respiratory infection clinic in a university hospital during the outbreak of COVID-19. Real-time polymerase chain reaction (RT-PCR) was performed, and the results of the two specimens were compared.ResultsTwo-hundred pairs of samples were collected. Sixty-nine (34.5%) individuals were male, and the median (interquartile) age was 36 (28–48) years. Using nasopharyngeal and throat swab RT-PCR as the reference standard, the prevalence of COVID-19 diagnosed by nasopharyngeal and throat swab RT-PCR was 9.5%. The sensitivity and specificity of the saliva sample RT-PCR were 84.2% (95% CI 60.4%–96.6%), and 98.9% (95% CI 96.1%–99.9%), respectively. An analysis of the agreement between the two specimens demonstrated 97.5% observed agreement (κ coefficient 0.851, 95% CI 0.723–0.979; p < 0.001).ConclusionsSaliva might be an alternative specimen for the diagnosis of COVID-19. The collection is non-invasive, and non-aerosol generating. This method could facilitate the diagnosis of the disease, given the simplicity of specimen collection and good diagnostic performance.     

Human papovavirus in a routine urine specimen of a four-year-old boy

Cellular changes produced by viruses can be readily identified using light microscopy and Papanicolaou stain of a fixed specimen. These findings can then be confirmed by viral culture and/or electron microscopy studies. Human polyomavirus, common in transplant recipients or otherwise immunocompromised patients, is one virus that can be identified using these methods. The following is a case study of a 4-yr-old boy with no known immune impairment who exhibited human papovavirus (polyomavirus) on a routine urine examination. The diagnosis was confirmed by electron microscopy.     

Evaluation of the core antigen assay as a second-line supplemental test for diagnosis of active hepatitis C virus infection

The British Columbia Center for Disease Control laboratory performs approximately 95% of all hepatitis C virus (HCV) antibody tests for the province's 4 million inhabitants. In 2002, the laboratory tested 96,000 specimens for anti-HCV antibodies, of which 4,800 (5%) were seroreactive and required confirmation of active infection. Although HCV RNA assays with a sensitivity of 50 IU/ml or less are recommended for the confirmation of active HCV infection, given the large number of seroreactive specimens tested annually, we evaluated the Ortho trak-C assay (OTCA) as a second-line confirmatory test and determined its limit of detection (LoD). Of 502 specimens from treatment-na?ve anti-HCV-positive individuals, 478 had sufficient volumes for evaluation by the OTCA and HCV RNA tests. Core antigen was not detected in 147 of 478 (30.8%) of these specimens, of which 37 of 147 (25.2%) were shown to be viremic by the VERSANT HCV (version 3.0) (branched-DNA) assay and/or the VERSANT HCV qualitative assay. Testing of 144 replicates of a World Health Organization standard dilution series indicated that the LoD of OTCA was approximately 27,000 IU/ml. This LoD is consistent with the inability of OTCA to detect core antigen in clinical specimens with low viral loads. We conclude that OTCA has limited value as a confirmatory test for the diagnosis of active HCV infection because 37 of 367 (10%) of viremic specimens had undetectable core antigen. Qualitative HCV RNA testing remains the present standard for the confirmation of active HCV infection in the diagnostic setting.     

A simple time-saving device for urine specimen preparation.

A technique is described for quick urine specimen preparation with the use of a simple device to save time and laboratory material.     

Dihydralazine hepatitis. Morphologic and clinical criteria for diagnosis

70 cases of acute dihydralazine-associated hepatitis with centrolobular or confluent necroses, registered in the files of the Berlin-Friedrichshain Institute of Pathology, between 1981 and 1985, were classified into 3 types of diagnostic probability for differential diagnosis versus virus hepatitis. Classification was conducted according to recommendations given by a working group of pathologists, specialised in liver pathology. 42 cases out of this material had come from Prenzlauer-Berg Hospital, Department of Infectious Diseases, and were re-examined under clinical aspects. 6 of them were discarded from evaluation. Type I proved to be of high diagnostic reliability, as was seen from 61% of all cases. Only 3 cases had to be discarded from that group and were associated with other drugs, such as halothane, methyldopa, and propranolol. The following clinical parameters proved to be of particular value for definite assessment of drug-induced hepatitis: time of exposure (for analysis of co-medication), time of recovery, and re-exposure test. Only circumstantial evidence so far can be provided for all histological types to causative relationship between drug ingestion and hepatitis. Compliance with mandatory notification should be ensured in all cases, since suspicious cases are explicitly included. Higher sex-related disposition of women to drug-induced hepatitis was confirmed in our material, with the female-to-male ratio being 3:1.     

Evaluation of rapid tests for diagnosis of acute hepatitis E

BackgroundHepatitis E virus diagnosis still presents difficulties due to discordant results among diagnostic tests.ObjectivesThe aim of this study was to evaluate the performance of two rapid tests for detection of anti-HEV IgM antibodies.Study designThe rapid tests were compared with three commercial anti-HEV ELISA assays and one Real-Time PCR assay on 59 sera from patients with acute viral non-AC hepatitis.ResultsThe presence of anti-HEV IgM antibodies was evaluated by two rapid tests (Wantai and Assure) on 25 HEV RNA positive samples. Anti-HEV IgM antibodies were detected in 24/25 and 23/25 samples respectively. The sensitivity and specificity of Wantai and Assure Rapid tests were evaluated using the 25 HEV RNA positive samples and 50 HEV RNA negative samples (including sera from acute-phase HAV and HBV infections and blood donors). Overall, the sensitivity of Wantai Rapid and Assure Rapid tests was 96.1% and 92.6% respectively; the specificity of the 2 tests was 100%.ConclusionOur data suggest the potential use of anti-HEV IgM rapid assays as a first line test in primary health care settings, particularly useful for patients with chronic liver disease or pregnant women who urgently need an antiviral treatment.     

Evaluation of a two-minute test for urine screening

A study was conducted to evaluate the ability of a urine filtration system (Bac-T-Screen, Marion Laboratories, Inc., Kansas City, Mo.) to detect negative urine cultures within 2 min. A total of 1,000 urine specimens were tested with the Bac-T-Screen and compared with a standard semiquantitative culture plate method and the Autobac system (General Diagnostics, Warner-Lambert Co., Morris Plains, N.J.). Of the 1,000 clean voided urine specimens tested, 246 specimens had colony counts greater than or equal to 10(5) CFU/ml by the culture plate method. Of these, the Bac-T-Screen detected 65.4% (161 of 246), and the Autobac detected 63.0% (155 of 246). When pure cultures of diphtheroids, lactobacilli, and viridans streptococci other than group D and cultures containing multiple organisms were considered to be contaminants and, therefore, were excluded, there were 106 pure cultures of probable pathogens of which the Bac-T-Screen detected 76.4% (81 of 106) and the Autobac detected 90.6% (96 of 106). Some 133 specimens were uninterpretable with the Bac-T-Screen because 36 clogged the filter and 97 left a residual pigment on the filter. A majority of those clogging the filter (69.4%) had positive plate counts, whereas the majority of the pigmented urines had negative plate counts. Of those urine specimens tested. 754 were negative by the culture plate method. The false-positive rates for Bac-T-Screen and Autobac were 16.2 and 5.8%, respectively. As a urine screen, the Bac-T-Screen has a negative predictive value comparable to the Autobac system and has the advantage of being a 2-min test.     

Monitoring human chorionic gonadotrophin level: evaluation of urine as an alternative specimen type

Although urine has been used widely for the qualitative detection of human chorionic gonadotrophin (hCG), serum is chosen conventionally for the serial quantification of the hormone to monitor trophoblastic activity. In response to requests from both clinicians and patients regarding the use of urine as an alternative specimen type, we designed this comparative study to evaluate the possibility, taking into account both laboratory technique and the distribution of hCG within different body fluids. Using the Access Chemiluminescent Immunoassay System, total beta-hCG was measured in serum and urine (n = 30) collected from patients hospitalised for first-trimester abnormal pregnancy. Results obtained with normalised urine (corrected with urinary creatinine) and serum total beta-hCG correlated well (r = 0.98, P < 0.001), and we concluded that urine could be used as an alternative specimen type for the serial quantitation of hCG to monitor trophoblastic activity. However, the assay used must detect the common beta 2 epitope.     

Detection of HEV antigen as a novel marker for the diagnosis of hepatitis E

Infection with hepatitis E virus (HEV) may be diagnosed by the presence of HEV RNA or anti-HEV antibodies. An enzyme immunoassay (EIA) was developed for the detection of antigen. Twenty-four monoclonal antibodies (mAbs) were produced. An indirect sandwich EIA was developed to detect HEV antigen using a combination of three mAbs as coating antibodies. Approximately 44.6% (33/74), 28.6% (50/175), and none (0/27) of sera positive for anti-HEV IgM alone, both anti-HEV IgM and IgG, and anti-HEV IgG alone also were positive for HEV antigen using this EIA. Forty-two HEV antibody-positive sera were tested for HEV RNA and antigen in parallel and the concordance was 81.0% (34/42). All PCR products were found to belong to HEV genotype 4. In order to evaluate the temporal relationship between HEV antigen positivity and HEV RNA, anti-HEV IgG and IgM, and ALT concentrations, macaques were infected with HEV genotypes 1 and 4 and serial samples were collected. The results showed that the antigen EIA can detect the capsid proteins of both genotypes. HEV antigen was detectable prior to ALT elevation and the appearance of anti-HEV antibodies in the infected monkeys and lasted for several weeks in all cases. HEV antigen became detectable in the serum at almost the same time as HEV RNA in feces but persisted for 4 weeks less than HEV RNA. This assay should be valuable for the diagnosis of acute hepatitis E, particularly in the window period prior to seroconversion to anti-HEV.     

血清乙型肝炎病毒表面抗原大蛋白检测的临床意义与评价

目的通过对乙型病毒性肝炎(乙肝)患者血清中乙肝表面抗原大蛋白(LHBs)、HBV DNA、乙肝前S1(PreS1)及传统的乙肝两对半的检测与平行比较研究,探讨LHBs用于临床诊断乙型肝炎的意义。方法检测385例乙肝患者的血清标本,LHBs、PreS1及乙肝两对半采用酶联免疫方法,HBV DNA检测采用实时荧光定量PCR法。结果307份HBV DNA阳性标本中LHBs的阳性率(86.97%)明显高于PreS1的阳性率(49.5%),两者差异有统计学意义(P<0.05);LHBs含量与HBV DNA拷贝数呈正相关性,r=0.935;HBeAg阴性标本中LHBs的检出率为76.92%,比较HBV DNA的检出率(67.95%)无统计学意义;PreS1的检出率(45.73%)则明显低于LHBs和HBV DNA。结论乙肝患者血清中LHBs表达与HBV DNA拷贝数呈正相关,两者的检出率与符合率均高于PreS1,LHBs检测可以用于反映乙肝患者病毒复制的血清免疫学指标。     

Cytopathological diagnosis of adult retinoblastoma in a vitrectomy specimen

Retinoblastoma (RB) is extremely rare in adults. We describe a case of RB diagnosed by cytology in a vitrectomy specimen of a 23‐year‐old patient who presented with diminished visual acuity and retinal detachment in the absence of a clinically‐visible mass. Cytological examination of the vitreous fluid showed clusters of loosely cohesive atypical cells with high nuclear to cytoplasmic ratio and “salt and pepper” chromatin pattern in a background of normal neuronal retinal cells. Nuclear molding was present as well as numerous apoptotic bodies. The cells were focally positive for epithelial markers and showed strong and diffuse positivity for neuroendocrine markers. Ki‐67 stained 90% of the “atypical cells” nuclei, in contrast to nonneoplastic retinal neuronal cells, which were negative for the marker. A diagnosis of RB was rendered, and subsequently was confirmed in the enucleation specimen. The cytological differential diagnosis is discussed as well as the role that cytology and immunohistochemistry can play in differentiating neoplastic cells from normal retinal cellular elements in vitreous fluid specimens. Diagn. Cytopathol. 2010. © 2009 Wiley‐Liss, Inc.