bluuebunny/crossref_metadata_2025 huggingface dataset:
Stability of dried blood spot specimens for detection of human immunodeficiency virus DNA by polymerase chain reaction
Authors: Cassol S , Salas T , Gill M J , Montpetit M , Rudnik J , Sy C T , O'Shaughnessy M V
Published: January 2020

Blood sampling on filter paper has many advantages for the detection of perinatal human immunodeficiency virus (HIV) infection by the polymerase chain reaction (PCR). However, if the method is to be widely used, an assessment of its performance under field conditions is required. To simulate conditions in the field, 50-microliters aliquots of whole blood containing low levels of HIV proviral DNA (4 to 1,024 copies per 100,000 nucleated cells) were spotted onto filter paper; dried; and subjected to heat, humidity, and prolonged storage at room temperature. After exposure, the DNA was recovered and amplified with primers to human leukocyte antigen DQ alpha- and HIV-specific sequences. Treatment at 37 degrees C and 60% humidity for 7 days, storage for 12 weeks at 22 degrees C, and freeze-thawing twice had no adverse effect on PCR reactivity when compared with the results obtained with reference spots stored at -20 degrees C. The lower limits of HIV detection in all tests ranged from 4 to 16 HIV copies per 100,000 cells. Fixation in 70% ethanol improved the amplification of low levels of HIV DNA and reduced biohazard risks. These findings suggest that dried blood spots will provide a powerful new resource for testing for HIV by PCR, especially in remote areas where refrigeration and immediate sample processing are unavailable.
Differentiation of Moraxella nonliquefaciens, M. lacunata, and M. bovis by using multilocus enzyme electrophoresis and hybridization with pilin-specific DNA probes
Authors: Tønjum T , Caugant D A , Bøvre K
Published: January 2020

Genetic relationships among strains of Moraxella nonliquefaciens, M. lacunata, and M. bovis were studied by using multilocus enzyme electrophoresis and DNA-DNA hybridization. The 74 isolates analyzed for electrophoretic variation at 12 enzyme loci were assigned to 59 multilocus genotypes. The multilocus genotypes were grouped in four major clusters, one representing strains of M. nonliquefaciens, two representing strains of M. lacunata, and one comprising strains of M. bovis and the single strain of M. equi analyzed. DNA-DNA hybridization with total genomic probes also revealed four major distinctive entities that corresponded to those identified by multilocus enzyme electrophoresis. The two distinct clusters recognized among the M. lacunata strains apparently corresponded to the species previously designated M. lacunata and M. liquefaciens. Distinction of the four entities was improved by hybridization with polymerase chain reaction products of nonconserved parts of pilin genes as DNA probes. With these polymerase chain reaction probes, new isolates of M. nonliquefaciens, M. lacunata, M. liquefaciens, and M. bovis can be identified easily by hybridization.
Comparison of commercial serological tests for detection of Helicobacter pylori antibodies
Authors: Talley N J , Kost L , Haddad A , Zinsmeister A R
Published: January 2020

Serological testing for Helicobacter pylori infection is one of the diagnostic methods of choice. Various commercial kits that use different antigens have been developed, but data on their diagnostic accuracy and direct comparisons between the tests are lacking. We aimed to evaluate the sensitivity, specificity, and predictive value of three immunoglobulin G enzyme-linked immunosorbent assay kits: Pylori stat, Helico-G, and Premier H. pylori. Serum samples and gastric biopsy findings from 76 patients were evaluated. We found by using a priori cutoff values that the Pylori stat, Helico-G, and Premier kits had overall sensitivities of 96, 96, and 88%, respectively, and specificities of 94, 86, and 96%, respectively, compared with gastric biopsy findings. For 232 serum samples, the Pylori stat test and a previously validated standard serological assay on which the test was based disagreed in 3% of the cases, while for 76 samples that were tested, Helico-G and a previously validated standard assay on which it was based disagreed in 8% of the cases. The intra- and interassay precision of each of the test kits was high. We conclude that serology based on any of these commercial tests represents a reliable and valid method for the diagnosis of H. pylori whether or not highly purified antigens are used.
Coamplified positive control detects inhibition of polymerase chain reactions
Authors: Cone R W , Hobson A C , Huang M L
Published: January 2020

Polymerase chain reactions (PCR) may fail to react because the substrate DNA is absent (true negative) or because of inhibition of specific amplification (false negative). The use of positive controls can increase confidence in negative PCR results by ruling out failure due to inhibition as a cause of the lack of amplification products. This report describes the construction and application of coamplified positive controls for herpes simplex virus and human herpesvirus 6 amplifications. Herpes simplex virus and human herpesvirus 6 PCR products were modified to generate control PCR products in which the original probe sequences were replaced by a Drosophila probe sequence. Implementation of the coamplified controls increased our specimen throughput in comparison with the parallel control amplifications used previously. Clinical laboratories using PCR for diagnosis of infectious diseases may find positive controls particularly helpful for increasing confidence that negative amplifications represent truly negative specimens.
The magnetic immuno polymerase chain reaction assay for direct detection of salmonellae in fecal samples
Authors: Widjojoatmodjo M N , Fluit A C , Torensma R , Verdonk G P , Verhoef J
Published: January 2020

Direct polymerase chain reaction (PCR)-based detection with fecal specimens is hampered by inhibitory compounds, such as bilirubin and bile salts. These fecal compounds showed significant inhibition of PCR at low concentrations (10 to 50 micrograms/ml). For direct PCR analysis, fecal samples must be diluted 500-fold to overcome inhibition. Therefore, the magnetic immuno PCR assay (MIPA), which combines immunomagnetic separation by using specific monoclonal antibodies and PCR, was used to directly detect salmonellae in feces from humans. Immunomagnetically extracted stool samples needed to be diluted only 10-fold when 1 microgram of T4 gene 32 protein was added to the PCR. The MIPA sensitivity obtained was 10(5) CFU/ml of feces. A panel of monoclonal antibodies specific for Salmonella serogroups A to E was used to extract salmonellae from clinical samples. MIPA detection of salmonellae occurred with 11 out of 14 stool samples stored at 4 degrees C for 2 months. MIPA detection of salmonellae in stool samples is a promising, fast method for detection and identification.
Application of rejection criteria for stool ovum and parasite examinations
Authors: Morris A J , Wilson M L , Reller L B
Published: January 2020

We retrospectively determined the yield of 2,015 stool ovum and parasite (O&P) examinations performed over an 11-month period. Two aspects were evaluated: the yield of positive results from multiple specimens per patient versus the result of a single examination, and the yield of positive results from stools submitted after 3 days of hospitalization. There were 131 (6.7%) positive O&P examinations from 75 patients: for 35 patients the single examination was positive; for 18, two of two examinations were positive; and for 15, three of three examinations were positive. The remaining seven patients had at least one negative examination in a series, but pathogenic intestinal parasites were detected in only three of these patients. Seventeen patients with positive O&P examinations were inpatients: seven of nine patients examined for O&P within 3 days of admission had stool specimens that contained recognized pathogens, in contrast to only two of eight patients examined after 3 days of hospitalization. After reviewing the data and informing hospital clinics, wards, and physicians, we instituted the following policy for screening stool specimens submitted for O&P examination. Only one O&P examination was performed for each outpatient visit and for inpatients hospitalized for 3 days or less, and examinations were not performed on stools submitted after 3 days of hospitalization unless the clinician arranged for the examination on the basis of clinical need. Over the 6-month follow-up period, 29.9% of O&P requests were rejected, 22% for patients in hospital for longer than 3 days and 7.9% for multiple O&P requests. Of 265 initially rejected specimens, 22 (8.3%) were examined after the referring physician contacted the laboratory. None of these specimens was positive. We conclude that eliminating O&P examinations of patients hospitalized for more than 3 days and initially performing only one examination per patient significantly reduces the number of examinations performed and reduces laboratory charges without adversely affecting patient care.
Nosocomial transmission of rotavirus from patients admitted with diarrhea
Authors: Gaggero A , Avendaño L F , Fernández J , Spencer E
Published: January 2020

We studied the transmission of rotavirus (RV) in 950 patients under 2 years of age hospitalized for diarrhea in Santiago, Chile. Stool samples were collected every other day from all patients during their entire hospital stay. To trace nosocomial transmission, we mapped the ward at the time of detection of RV. Comparative study by polyacrylamide gel electrophoresis of 315 RV isolates (180 detected upon admission of patients and 135 attributed to nosocomial transmission) allowed the identification of 18 different electropherotypes. An electropherotype similar to that of a community-acquired case was found in the same room in 81% of nosocomial cases and in the ward in 92% of nosocomial cases. It was concluded that the infants admitted shedding RV are the major source of nosocomial transmission and there was not a RV strain that was particularly transmissible.
Serodiagnosis of Lyme borreliosis by western immunoblot: reactivity of various significant antibodies against Borrelia burgdorferi
Authors: Ma B , Christen B , Leung D , Vigo-Pelfrey C
Published: January 2020

The significance of various antibodies against Borrelia burgdorferi was studied by Western blot (immunoblot) by using 578 human serum samples. The proteins regularly detected by using samples from patients with Lyme borreliosis were those with bands with molecular masses of 94, 83, 75, 66, 60, 55, 46, 41, 39, 34, 31, 29, 22, and 17 kDa. The detectable frequencies of most of these proteins appeared to be significantly different between the sera from patients with Lyme borreliosis and those from normal control individuals as well as from the group with syphilis. The 39-kDa protein band recognized by polyvalent antibody was found to be the most significant marker for Lyme borreliosis. Furthermore, an anti-39-kDa immunoglobulin M response was detected in the samples from patients with early-stage Lyme borreliosis. Results from the use of monoclonal antibodies and patient sera revealed that the 39- and 41-kDa proteins may be structurally related but are immunologically distinct antigens. The significance of antibody reactivities to the 41-, 94-, 22-, 31-, and 34-kDa protein bands is also discussed.
Antigenemia in paracoccidioidomycosis
Authors: Freitas-da-Silva G , Roque-Barreira M C
Published: January 2020

Severe forms of paracoccidioidomycosis (Pcm) are accompanied by intense immunological involvement characterized by depression of the cell-mediated immune response and by high levels of antibodies in serum with no protective function. These changes can be reversed by antifungal treatment. It has been suggested that antigens of Paracoccidioides brasiliensis released into the circulation during the active phase of the disease may be involved in the genesis of the changes in the immune response. In the present study, we evaluated the antigenemia of patients with Pcm using a competitive enzyme-linked immunosorbent assay (ELISA-c) capable of detecting 6 ng of antigen per ml of serum. Twenty-seven of 88 serum samples tested gave positive results, with the highest frequency of positivity being detected in patients with the severe acute form of the disease; these patients had the highest antigen levels (0.03 to 3.4 micrograms/ml). Follow-up of one case showed a correlation between antigen levels in serum and evolution of the disease. False-positive reactions were observed in sera from patients with histoplasmosis, aspergillosis, and cryptococcosis. The results indicate that the described method has potential for clinical application, especially with respect to the evaluation of disease activity. Quantification of fungal antigens in the serum of patients with active Pcm represents an objective parameter for the study of the physiopathology of the disease.
Cross-infection risks associated with current procedures for using high-speed dental handpieces
Authors: Lewis D L , Boe R K
Published: January 2020

When a dye solution used to simulate patient material was either injected into high-speed dental handpiece (drill) waterlines or applied to the equipment externally, internal air turbine chambers became contaminated. These chambers served as a reservoir of the material, which was slowly dislodged by air expelled during subsequent handpiece operation and which was diluted by water spray used for cooling the drilling surface. Considering the fact that patient materials could reside in internal parts of the equipment that are not usually disinfected and that the material may be subsequently sprayed into cuts and abrasions in the oral cavity, the common approach to reprocessing handpieces (external wiping in combination with flushing) may pose unacceptably high risks to those individuals treated soon after infected patients. Therefore, unless reliable data on cross-infection frequencies are obtained and prove it unnecessary, heat-treating high-speed handpieces between each patient should be considered an essential component of standard procedures whenever universal precautions are practiced in dentistry.
Development of a polymerase chain reaction-based diagnosis of Trichomonas vaginalis
Authors: Riley D E , Roberts M C , Takayama T , Krieger J N
Published: January 2020

We developed a polymerase chain reaction (PCR)-based test for detecting the protozoan parasite Trichomonas vaginalis. Genomic libraries were constructed from two independent clinical isolates of T. vaginalis. From these libraries, 12 genomic clones were purified, sequenced, and then screened for uniqueness by computer-assisted sequence comparisons. PCR reactions were performed to evaluate eight PCR-primer pairs, including a primer pair that targeted the T. vaginalis ferredoxin gene. All eight primer pairs yielded PCR products of the expected sizes. However, six of the primer pairs amplified their respective target sequences in limited numbers of clinical T. vaginalis isolates, suggesting the presence of significant genomic variability among isolates. An exception was a primer pair, termed TVA5-TVA6, that amplified a 102-bp genomic sequence, termed A6p, in all of 24 clinical isolates. The A6p sequence was not detected by PCR in human DNA or in a wide variety of flagellates, ciliates, or bacteria tested. The A6p sequence appears highly selective for a broad range of T. vaginalis isolates and holds promise for PCR-based diagnosis of the parasite.
Comparison of sensitivities of three commercial MRC-5 cell lines grown in shell vials to cytomegalovirus and responses to enhancing agents
Authors: West P G , Hartwig R A , Baker W W
Published: January 2020

MRC-5 cells grown in shell vials from three suppliers (Whittaker Bioproducts, Bartels, and ViroMed) were evaluated for sensitivity to human cytomegalovirus. Previous studies done in our laboratory showed that treatment of the cells with dexamethasone (DEX) and dimethyl sulfoxide (DMSO) or these agents plus calcium improved the sensitivity of cells from Whittaker. In this study, we compared the response of the Whittaker cells with those of the cells from Bartels and ViroMed, both treated and untreated, when tested with strain AD169, a strain of wild-type virus, and 41 clinical specimens from various sources. A progressive improvement in sensitivity to AD169 and the wild-type virus was seen for all cell lines with DEX-DSMO and DEX-DMSO-calcium treatments, although even untreated, the Whittaker cells were much more sensitive. Detection of most of the positive clinical specimens was enhanced by both treatments with the Whittaker cells. DEX-DMSO had little or no effect on Bartels or ViroMed cells, while DEX-DMSO-calcium usually effected some increase in the number of inclusions. Six specimens were recovered only on Whittaker cells. Protein studies were done on all groups to determine the effect, if any, of the treatments on the metabolism of the cells.
Modified indirect hemagglutination test for detection of treponemal antibodies in finger-prick blood
Authors: Backhouse J L , Lee M H , Nesteroff S I , Hudson B J , Hamilton P A
Published: January 2020

A modified indirect hemagglutination test for the detection of treponemal antibodies was developed for use with finger-prick blood. By using paired serum and absorbed finger-prick blood from 58 patients from an area previously endemic for yaws and 12 patients without yaws, the modified hemagglutination test was compared with a hemagglutination test for Treponema pallidum and the fluorescent treponemal antibody absorption test. The modified hemagglutination test showed 100% specificity and an overall agreement of 96.5% with the hemagglutination test for T. pallidum and 94.8% with the fluorescent treponemal antibody test. The modified hemagglutination test appears to be a simple and economical test that is suitable for use in large epidemiological surveys for yaws.
Diverse populations of Legionella pneumophila present in the water of geographically clustered institutions served by the same water reservoir
Authors: Bezanson G , Burbridge S , Haldane D , Yoell C , Marrie T
Published: January 2020

We cultured potable water from seven institutions (six hospitals and one medical school) every 2 weeks for 6 months for Legionella pneumophila. All of the institutions were located close to each other and received water from the same freshwater source. Two institutions (the medical school and hospital F, a maternity hospital) never had L. pneumophila isolated from their potable water. The remaining five had 17 to 72% of their water samples positive for L. pneumophila. Most of the isolates were serogroup 1; however, in hospital B serogroup 5 accounted for 56% of the isolates. Oxford and OLDA monoclonal antibody subtypes of L. pneumophila serogroup 1 coexisted in four of the five institutions, while subtype France only was found in one institution. All 10 isolates from this institution lacked plasmids. The other four institutions had Legionella populations with plasmid profiles II, III, and VI. Two of these institutions also had isolates with no plasmids. The distribution of the plasmid types was significantly different for all institutions except C and D. The distribution of monoclonal antibody subtypes was significantly different for L. pneumophila isolates recovered from institutions C and D. There were no characteristics that distinguished the culture-positive institutions from the culture-negative areas. We conclude that diverse populations of L. pneumophila exist within these institutions despite their geographic proximity and identical potable water source.
Streptococcus pyogenes pharyngitis: characterization of strains by multilocus enzyme genotype, M and T protein serotype, and pyrogenic exotoxin gene probing
Authors: Musser J M , Gray B M , Schlievert P M , Pichichero M E
Published: January 2020

Multilocus enzyme electrophoresis, serological characterization of M and T proteins, and probing for pyrogenic exotoxin A and C genes were used to investigate the bacteriologic epidemiology of strains of Streptococcus pyogenes recovered primarily from patients with recurrent pharyngitis. A total of 164 strains recovered from individuals living in nine states of the United States was analyzed. Two-thirds of the patients in our sample were infected with the homologous strain following antibiotic therapy and presumably represented treatment failures, whereas the other one-third of the patients were infected with a heterologous strain after therapy and probably represented reinfections. Multilocus enzyme electrophoresis was as efficacious in strain discrimination as serologic typing techniques were and, in addition, successfully characterized all organisms that were serologically nontypeable. Two clones of S. pyogenes responsible for most of the episodes of toxic shock-like syndrome in the United States are geographically widespread, but they vary by locality in the frequency of their occurrence. Compared with a sample of strains cultured from patients whose pharyngeal infections were eliminated by antimicrobial therapy, these two clones were statistically overrepresented among organisms that cause recurrent pharyngitis.
Multicenter evaluation of five commercial rubella virus immunoglobulin G kits which report in international units per milliliter
Authors: Dimech W , Bettoli A , Eckert D , Francis B , Hamblin J , Kerr T , Ryan C , Skurrie I
Published: January 2020

In a multicenter study, the consistency of international units expressed by five commercially available rubella virus immunoglobulin G kits was evaluated. The linearity and within-run and between-run precision were determined for each kit. All kits demonstrated good linearity and had within-run and between-run precision coefficients of variation ranging from 5.1 to 21.7% and from 9.5 to 51.0%, respectively. To compare the international units expressed, the results from 40 samples tested in duplicate were compared with the results of a reference enzyme immunoassay calibrated with World Health Organization international standard serum and a hemagglutination inhibition test. The results of the kits were plotted against those of the reference tests, and linear regression analysis was applied. The Pearson correlation coefficient ranged from 0.64 to 0.75 when the commercial kit results were compared with those of the reference enzyme immunoassay, indicating only a moderate degree of correlation. Therefore, the international units expressed by the commercial kits are insufficiently consistent to be of practical use in diagnostic clinical microbiology.
Evaluation of an automated immunodiagnostic assay system for direct detection of herpes simplex virus antigen in clinical specimens
Authors: Johnston S L , Hamilton S , Bindra R , Hursh D A , Gleaves C A
Published: January 2020

The Vitek ImmunoDiagnostic Assay System (VIDAS) is a 2 1/3-h automated qualitative enzyme-linked fluorescent immunoassay developed for the direct detection of herpes simplex virus (HSV) antigen in clinical specimens. A total of 356 clinical specimens submitted for HSV isolation were prospectively evaluated with the VIDAS, and the results of the technique were compared with those of both HSV isolation in cell culture and Herpchek, a nonautomated enzyme immunoassay. Compared to cell culture, VIDAS had a sensitivity of 91.6% and a specificity of 89.3%, with positive and negative predictive values of 82.6 and 95.0%, respectively. In comparison to Herpchek, VIDAS had a sensitivity of 93.7% and a specificity of 93.0%, with positive and negative predictive values of 89.4 and 95.9%, respectively. The results demonstrated that the VIDAS required minimal manipulation in order to produce results comparable to those of Herpchek and HSV isolation in cell culture.
Comparison of plasmid profiles, phage types, and antimicrobial resistance patterns of Salmonella enteritidis isolates in the United States
Authors: Rodrigue D C , Cameron D N , Puhr N D , Brenner F W , St Louis M E , Wachsmuth I K , Tauxe R V
Published: January 2020

To evaluate the laboratory techniques for subtyping isolates of Salmonella enteritidis, we compared the plasmid profiles (PP), phage types (PT), and antimicrobial susceptibility patterns (AS) of two nationally representative samples of sporadic human S. enteritidis isolates from 1979 (n = 28) and 1984 (n = 37), 43 isolates from 20 outbreaks of S. enteritidis infections between 1983 and 1987, and 46 animal isolates selected from the U.S. Department of Agriculture Veterinary Services Laboratory in 1986 and 1987. Sporadic and outbreak isolates from humans showed similar rates of resistance to at least one of a panel of antimicrobial drugs (23 and 14%, respectively), PT (91 and 98%, respectively), and PP (97 and 100%, respectively). Sixteen different PP were identified in sporadic, outbreak, and animal isolates; two PP accounted for 76% of sporadic and outbreak isolates. Sporadic human isolates were of PT 8 (42%), of PT 13a (37%), nontypeable (9%), of PT 14b (8%), of PT 9a (3%), and of PT 13 (2%). Outbreak human isolates had similar distributions of PT. PT 8 was associated with poultry: 58% (7 of 12) of the poultry isolates but only 24% (8 of 34) of the isolates from other animals were of PT 8 (P less than 0.04). Although antimicrobial susceptibility patterns do not appear as useful as an epidemiologic marker, PP and PT effectively subtyped S. enteritidis.
Comparison of MB-Check, BACTEC, and egg-based media for recovery of mycobacteria
Authors: Abe C , Hosojima S , Fukasawa Y , Kazumi Y , Takahashi M , Hirano K , Mori T
Published: January 2020

The rate of recovery and time to the detection of mycobacteria from clinical specimens were measured for biphasic (MB-Check; Nippon Roche Co., Ltd., Tokyo, Japan) and radiometric (BACTEC; Nippon Becton Dickinson Co., Ltd., Tokyo, Japan) liquid-based culture systems and egg-based media (3% Ogawa and Ogawa K). From the 245 sputum specimens processed, a total of 86 (35.1%) mycobacterial isolates were detected. Of these, 81 (94.2%) and 80 (93.0%) isolates were detected with the MB-Check and BACTEC systems, respectively, and 65 (75.6%) isolates were detected with the 3% Ogawa egg method. The difference in the percentages of positive cultures between the two systems based on liquid media and the 3% Ogawa egg method was significant (P less than 0.01). This difference was even greater among smear-negative specimens. The detection time was shorter with the liquid-based systems. The mean times to the detection of the Mycobacterium tuberculosis complex were 19.1 days with the MB-Check system, 13.4 days with the BACTEC system, and 21.7 days with the 3% Ogawa egg method. These results indicate that both the MB-Check and the BACTEC systems, based on liquid media, are efficient for the recovery of mycobacteria.
Multicenter evaluation of the use of Haemophilus test medium for broth microdilution antimicrobial susceptibility testing of Streptococcus pneumoniae and development of quality control limits
Authors: Jorgensen J H , Doern G V , Ferraro M J , Knapp C C , Swenson J M , Washington J A
Published: January 2020

A five-laboratory collaborative study was undertaken to determine the precision and accuracy of broth microdilution susceptibility tests of Streptococcus pneumoniae isolates performed with Haemophilus test medium (HTM) compared with tests performed with lysed horse blood-supplemented Mueller-Hinton broth (LHB). The intra-and interlaboratory reproducibilities of MICs of 10 antimicrobial agents determined with the two media were found to be quite similar and highly reproducible in both media. On the basis of favorable performance in this study, S. pneumoniae ATCC 49619 is recommended as a quality control strain to assess the performance of HTM when this medium is used for testing of pneumococci. Testing of 293 unique clinical isolates of S. pneumoniae with both media in the respective participant laboratories allowed a direct comparison of MIC results and a calculation of interpretive error rates. Although there were some slight differences between MICs determined with HTM and MICs determined with LHB, few very major or major errors resulted from testing the clinical isolates against the 10 antimicrobial agents. However, MIC-interpretive criteria specific for S. pneumoniae should be developed and promulgated through a national consensus mechanism.
Comparison of a one-step and a two-step polymerase chain reaction with degenerate general primers in a population-based study of human papillomavirus infection in young Swedish women
Authors: Evander M , Edlund K , Bodén E , Gustafsson A , Jonsson M , Karlsson R , Rylander E , Wadell G
Published: January 2020

The prevalence of human papillomavirus (HPV) infection in cervical cell scrapes from young women was determined by polymerase chain reaction (PCR) by using general primer pairs localized within the L1 region. With a one-step general PCR, 5.9% (35 of 590) of young women in a population-based study were found to contain HPV DNA. The proportion of HPV-positive women increased with age, from 1.4% (1 of 69) among women aged 19 years to 9.2% (13 of 142) among women aged 25 years. Among the cervical scrapes from women with normal cytology, 5.6% (30 of 539) harbored HPV DNA. A total of 5 of 19 (26.3%) of the women with pathological signs were positive for HPV DNA. By a two-step PCR, using nested general primers, 20.3% (118 of 581) of all women were shown to contain HPV DNA. The proportion of HPV-positive women also increased with age, from 17.4% (12 of 69) among women aged 19 years to 31.9% (43 of 135) among women aged 25 years, when the two-step PCR was used. Some 19.2% (102 of 530) of cervical scrapes from women with normal cytology contained HPV DNA. Among the women with pathological signs, 16 of 19 (84.2%) were positive for HPV DNA. The HPV DNA-positive specimens were demonstrated to contain HPV type 6, 11, 16, 18, 31, 33, 35, 39, 40, 45, 55, or 56. The most prevalent HPV types were 6 (2.0%) and 16 (2.7%). More than one type was found in 16 specimens. Sixty HPV-positive samples could not be typed.
Isolation of Corynebacterium group D2 from two dogs with urinary tract infections
Authors: Elad D , Aizenberg I , Shamir M , Soriano F , Shlomovitz S
Published: January 2020

Corynebacterium group D2 was isolated from two dogs with urinary tract infections. The isolates were resistant in vitro to all tested antibacterial drugs except vancomycin. One dog was successfully treated with this antibiotic, while the other died before treatment could be initiated.
Indirect hemagglutination assay for diagnosis of Escherichia coli O157 infection in patients with hemolytic-uremic syndrome
Authors: Bitzan M , Karch H
Published: January 2020

An indirect hemagglutination assay consisting of sheep erythrocytes coated with lipopolysaccharide (LPS) from Shiga-like toxin-producing Escherichia coli O157 was used for the serological diagnosis of E. coli O157 infections in children with classical (enteropathic) hemolytic-uremic syndrome (HUS). One week after the onset of diarrhea (acute phase of the disease), the E. coli O157 antibody titer was greater than or equal to 1:4,096 in 22 of 27 patients with HUS, compared with 4 of 249 controls, the majority of whom had O157 antibody titers of between 1:4 and 1:256. This antibody response was observed in HUS patients with stool cultures positive and negative for E. coli O157. Selective absorption with homologous LPS and heterologous LPS showed that the antibody response was specific for E. coli O157. Because of its simplicity and ease of interpretation, the indirect hemagglutination assay described in this paper is recommended for the serological diagnosis of E. coli O157 infections in patients with HUS.
Stability of human immunodeficiency virus type 1 antibodies in whole blood dried on filter paper and stored under various tropical conditions in Kinshasa, Zaire
Authors: Behets F , Kashamuka M , Pappaioanou M , Green T A , Ryder R W , Batter V , George J R , Hannon W H , Quinn T C
Published: January 2020

The use of whole-blood spots on filter paper for the detection of antibody to human immunodeficiency virus type 1 (HIV-1) was evaluated during a 20-week period under a variety of storage environments simulating the harsh tropical field conditions in Kinshasa, Zaire. During the first 6 weeks of storage, all replicates of high- and low-titer HIV-1-positive reference samples remained positive by enzyme immunoassay and Western blotting (immunoblotting), and all replicates of HIV-1-negative samples remained negative under all storage conditions. However, hot and humid storage conditions for up to 20 weeks caused a progressive decline in enzyme immunoassay optical density ratio values, which was particularly noticeable in samples with a low HIV-1 antibody titer. Harsh tropical operational conditions did not cause any repeatedly false-positive results during the 20-week storage period. The use of gas-impermeable bags with desiccant for the storage of blood spots on filter paper improved the stability of HIV-1 antibody detection over time and is recommended for the storage of whole-blood spots on filter paper in harsh tropical field settings.
Protein composition of the hepatitis B virus e antigen in the natural course of disease and following interferon therapy
Authors: Campillo M L , Quiroga J A , Bartolomé J , Moraleda G , Castillo I , Carreño V
Published: January 2020

The protein composition of hepatitis B virus (HBV) e antigen (HBeAg) in serum was analyzed for 63 viremic patients with chronic HBV and found to consist of polypeptides with molecular masses of 16, 18, and 20 kDa (P16e, P18e, and P20e, respectively). Several experiments demonstrated their viral nature and HBeAg specificity and that P16e occurs either in free soluble form or as aggregates or immunocomplexes, while P18e and P20e occur essentially as immunocomplexes. Of 63 patients, 45 (71%) had P16e, P18e, and P20e, and the remaining 18 (29%) had only P16e. During the natural history of the disease, spontaneous clearance of HBV DNA and HBeAg took place only in six of nine (67%) patients with the three HBe polypeptides but in none of the five patients having P16e alone (P less than 0.05). Similarly, 22 of 23 (96%) patients responding to interferon therapy had P16e, P18e, and P20e, but these polypeptides occurred in only 14 of 26 (54%) nonresponder patients (P less than 0.001). Following the loss of HBV DNA and HBeAg, before the development of anti-HBe, the only HBe species detected were P18e and P20e, but these became no longer detectable after complete normalization of liver function tests. Therefore, persistence of P16e represents a failure to recover from HBV, and the appearance of HBe polypeptides P18e and P20e is associated with virus clearance and a favorable outcome of the disease.
Age-related interference with Chlamydia pneumoniae microimmunofluorescence serology due to circulating rheumatoid factor
Authors: Verkooyen R P , Hazenberg M A , Van Haaren G H , Van Den Bosch J M , Snijder R J , Van Helden H P , Verbrugh H A
Published: January 2020

Microimmunofluorescence (MIF) serology is commonly used in the diagnosis of chlamydial infections. In the MIF assay, Chlamydia pneumoniae elementary bodies were used to detect C. pneumoniae immunoglobulin G (IgG) and IgM antibodies in paired serum samples from 286 patients with respiratory illnesses. In 69 patients, MIF serology was compared with C. pneumoniae cultures. All C. pneumoniae cultures remained negative. However, 205 (71%) of 286 patients were C. pneumoniae antibody positive and 64 (22%) had MIF test results indicating recent infection; 11 showed a fourfold increase in IgG titer, 18 had IgG titers of greater than or equal to 1:512, and 41 had IgM titers of greater than or equal to 1:16. In 35 (55%) of 64 patients, a recent-infection diagnosis was based on C. pneumoniae IgM antibodies only. However, 78% of C. pneumoniae IgM-positive patients had circulating rheumatoid factor (RF) by rheumatoid arthritis latex assay. RF positivity increased with age. After absorption with anti-human IgG, all C. pneumoniae IgM-positive sera became C. pneumoniae IgM negative in the MIF assay. Twenty-five patients with active rheumatoid arthritis but without respiratory illness were also tested; 14 were C. pneumoniae IgG positive and C. pneumoniae IgM positive as well. Absorption of IgG from these RF-containing sera invariably resulted in disappearance of reactivity in the MIF IgM assay. We conclude that with age the serologic diagnosis of recent C. pneumoniae infection becomes increasingly prone to false-positive results unless sera are routinely absorbed prior to MIF IgM testing.
Comparison of the Western immunoblot assay and a glycoprotein G enzyme immunoassay for detection of serum antibodies to herpes simplex virus type 2 in patients with AIDS
Authors: Safrin S , Arvin A , Mills J , Ashley R
Published: January 2020

Herpes simplex virus type 2 (HSV-2) seroprevalence in 68 patients with AIDS was 77% by Western blot (immunoblot) and 44% by glycoprotein G-2 immunoassay. Each of 16 patients with culture-proven HSV-2 infection was positive by Western blot versus 8 by glycoprotein G-2 immunoassay. No differences in age, race, duration of AIDS, acyclovir usage, or HSV-1 seroprevalence were found to explain differences in sensitivity.
Assay of urease-inhibiting activity in serum from children infected with Helicobacter pylori
Authors: Thomas J E , Whatmore A M , Kehoe M A , Skillen A W , Barer M R
Published: January 2020

In order to provide a basis for obtaining further information concerning the host response to Helicobacter pylori urease, four assay methods for detecting urease-inhibiting activity in serum were examined. A quantitative assay, established in a COBAS BIO centrifugal fast analyzer and based on detection of the consumption of NADH by glutamate dehydrogenase stimulated by ammonia production, was considered most suitable for large-scale serological work. Serum samples from 63 children (aged 5 to 16 years), 28 of whom had seropositive H. pylori gastritis, were assayed. One of the serum samples in this latter group showed significant inhibitory activity. This serum sample was one of 13 in the seropositive group known to bind to urease antigen. It showed no inhibitory activity against Bacillus pasteurii or jack bean urease. Protein A binding and heat treatment indicated that the inhibitory activity was immunoglobulin G mediated. The patient from whom this sample was collected showed no distinctive features in his illness. The COBAS BIO analyzer-based urease inhibition assay provides a new tool for studying one aspect of the host response to H. pylori infection.
Niacin-positive Mycobacterium kansasii isolated from immunocompromised patients
Authors: Nachamkin I , MacGregor R R , Staneck J L , Tsang A Y , Denner J C , Willner M , Barbagallo S
Published: January 2020

Niacin-positive Mycobacterium kansasii was isolated from three patients, two with respiratory infections and one with a perirectal abscess. The isolates were phenotypically similar to other strains of M. kansasii, differing only in their ability to produce niacin. This phenotype has been reported only twice in the literature, during the 1960s.
Fluorogenic substrates for differentiation of gram-negative nonfermentative and oxidase-positive fermentative bacteria
Authors: Kämpfer P , Kulies I , Dott W
Published: January 2020

A total of 803 strains of gram-negative nonfermentative and oxidase-positive fermentative bacteria (38 taxa) were investigated for their ability to hydrolyze 53 different fluorogenic 4-methylumbelliferyl- and beta-naphthylamide-linked substrates within 6 h of incubation. The hydrolysis of 16 fluorogenic substrates showed high separation index values among the tested taxa, was reproducible, and showed good agreement with data in the literature. In combination with other biochemical tests (like carbon substrate utilization tests and classical biochemical tests), hydrolysis profiles can improve the differentiation of gram-negative nonfermentative and oxidase-positive fermentative bacteria.
Optimal survival of Helicobacter pylori under various transport conditions
Authors: Soltesz V , Zeeberg B , Wadström T
Published: January 2020

The ability of clinical isolates and type strains of Helicobacter pylori to survive in Stuart transport medium, isotonic saline solution, and urea-containing isotonic saline was evaluated. The influences of temperature (4, 10, 15, 20, and 30 degrees C) and holding time (6 to 48 h) and the effect of exposure to air on survival were also studied. The recovery rate of H. pylori was highest from Stuart transport medium in comparison with the recoveries from the other transport media tested. We found that at holding temperatures above 15 degrees C the organisms became noncultivable within 6 h or less, while they survived for 2 days or longer at 10 degrees C. The presence of urea at a concentration of 2% (wt/vol) in isotonic saline resulted in the loss of viability of the organisms tested.
Length polymorphisms in tRNA intergenic spacers detected by using the polymerase chain reaction can distinguish streptococcal strains and species
Authors: McClelland M , Petersen C , Welsh J
Published: January 2020

Intergenic tRNA spacers from strains of streptococcal groups A, B, and G were amplified by using the polymerase chain reaction (PCR) at low stringency with consensus tRNA gene primers. Cloning and sequencing showed that many of the homologous intergenic spacers differed in length between species. The sequences of the tRNA genes that flank these polymorphic spacers were determined and used to synthesize fully complementary primers. With these primers at high stringency, PCR products which varied in lengths from 53 to 71 bp, depending on the species or strain, were obtained from streptococcal DNAs, even in the presence of a 1,000-fold mass excess of human DNA. PCR products, the lengths of which could also be used for classification, were obtained at high stringency from a few genera closely related to Streptococcus. No products were obtained from genomic DNAs from more distantly related genera. Production of species- or strain-specific tRNA intergenic length polymorphisms with primers that generate characteristic products from a variety of species within the same genus should be applicable to many organisms, including those that would otherwise be difficult to culture or identify.
Comparison of DNA fingerprints and somatic serotypes of serogroup B and E Pasteurella multocida isolates
Authors: Wilson M A , Rimler R B , Hoffman L J
Published: January 2020

The DNA fingerprint profiles and somatic serotypes of 71 Pasteurella multocida capsule serogroup B isolates, 13 capsule serogroup E isolates, and 16 somatic reference serotype strains were compared. Each of the 16 reference somatic serotypes had a unique DNA fingerprint profile with the HhaI restriction endonuclease. Fifty-four serogroup B isolates (isolated from classical cases of hemorrhagic septicemia) reacted with somatic serotype 2 or 5 antiserum and had DNA fingerprint profiles which resembled that of the serotype 2 reference strain. Seven DNA fingerprint profiles were found among 16 serogroup B strains representing other somatic serotypes. The DNA fingerprints of these isolates were different from the fingerprints of the 16 somatic reference serotype strains. All 13 serogroup E isolates had identical somatic serotypes and identical DNA fingerprint profiles when the HhaI endonuclease was used. The HhaI fingerprint profile of the serogroup E isolates did not match any fingerprint profile of the reference somatic serotype strains. Following DNA profiling with the HhaI endonuclease, the 13 serogroup E isolates were differentiated sequentially with HpaII restriction endonuclease. A descriptive identification epithet for P. multocida isolates was constructed. The descriptive epithet consists of serologic identification and sequential DNA profiles with restriction endonucleases HhaI and HpaII, respectively. DNA fingerprinting of P. multocida is a precise characterization method. In conjunction with serologic typing, it can further classify P. multocida isolates for epidemiologic studies.
Evaluation of the autoSCAN-W/A system for rapid (2-hour) identification of members of the family Enterobacteriaceae
Authors: O'Hara C M , Miller J M
Published: January 2020

We evaluated the ability of the Baxter autoSCAN-W/A System (MicroScan Division, Baxter Diagnostics, Inc., West Sacramento, Calif.) to use the rapid (2-h) gram-negative identification panel for accurate identification of members of the family Enterobacteriaceae. At 2 h, 353 of 467 (75.6%) strains in a challenge set of biochemically typical and atypical stock cultures were correctly identified to genus and species. Another 76 (16.3%) strains were correctly identified to genus and species after the performance of recommended additional biochemical testing. Thus, at 24 h, 91.9% of the 467 strains were correctly identified. Twenty-two strains (4.7%) were identified to the correct genus but the incorrect species, and 16 strains (3.4%) were misidentified. Of these 16 strains, 9 were incorrect at 2 h, and 7 were incorrect after the additional testing. Because the system is based on fluorogenic substrates, no conventional tests were readily available with which to compare aberrant reactions. These results suggest that the autoSCAN-W/A with its rapid gram-negative panels is acceptable for the identification of the Enterobacteriaceae in a clinical microbiology laboratory.
Statistical analysis of the incidence of positives in the examination of parasitological specimens
Authors: Mohr E , Mohr I
Published: January 2020

Results of the examination of 2,000 parasitological specimens were analyzed for differences between results obtained by a technologist working in parasitology only and those obtained by general microbiologists who were also performing other laboratory work. The relative share of specimens determined to be positive by the specialist was approximately twice as high as the relative share determined by the general microbiologists.
Evaluation of a competitive enzyme immunoassay for detection of Coxiella burnetii antibody in animal sera
Authors: Soliman A K , Botros B A , Watts D M
Published: January 2020

A competitive enzyme immunoassay (CEIA) was established and compared with other serological techniques for detecting Coxiella burnetii antibody in camels, goats, and sheep. This technique was evaluated because a conjugated anti-camel immunoglobulin was not available to serve as a direct signal for the demonstration of antigen-antibody reaction. A C. burnetii antibody-positive human serum and a peroxidase-conjugated anti-human immunoglobulin G were used as an indicator system competing against antibody in animal serum or as an indicator of the absence of antibody. Sera were considered antibody positive when the A414 of the test sera plus the competing positive antibody was less than or equal to 50% of the A414 of the negative-control serum plus the competing antibody. Antibody to C. burnetii was repeatedly demonstrated in 66% of camel serum samples (n = 200) by the CEIA. Among 48 camel serum samples, 71% were positive for antibody by CEIA versus 65% by EIA using peroxidase-labeled protein A. The CEIA detected C. burnetii antibody in 63% of sheep serum samples (n = 40) and in 50% of goat serum samples (n = 96), while the indirect fluorescent-antibody technique detected antibody in 38% of sheep and 34% of goat serum samples and the EIA detected antibody in 50% of sheep and 35% of goat serum samples. These data indicate that the CEIA is a reliable and sensitive technique for demonstrating C. burnetii antibody in camels, sheep, and goats.