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[Relationship of serum streptococcal antibodies to carrier state of beta-hemolytic streptococcus in throats of healthy schoolchildren].

The relationship of ASO, ASK and ASP to carrier state of beta-hemolytic streptococcus in throats of healthy schoolchildren was examined from September, 1980 to May, 1987. The responses to ASO significantly increased in children who detected group A, C and G streptococcus, and ASK in group A and G, and ASP in group A. The responses were especially high in children who detected group A, C and G streptococcus one after the other through the term of research, and in children detected another T type of streptococcus one after the other. Relatively high titers of ASO and ASK were obtained in sera of children from whom some groups or T types of streptococci were isolated several detection times. ASP titers were not concerned with these times. The elevation of antibodies in paired samples were found as follows; 7 cases in ASO, ASK and ASP together, 42 cases in ASO and ASK together, 3 cases in ASK and ASP together, 2 cases in ASO and ASP together, 41 cases ASO only, 38 cases in ASK only, 7 cases in ASP only. Among these cases with elevation of antibodies titers, several kinds of the change of streptococcal carrier state were observed between obtaining serum of twice. Especially 15 cases with change of carrier state from group A to group A were noticed. Among these 15 cases, 11 cases isolated another T type strain through the term of research, or found to contain the research-time when no streptococcal strain was detected. Both long term carrier state of the same group or the same type of streptococcal strain and change of carrier state of streptococcal group or T type lead the elevation of ASO and ASK titers.

Antibodies, Bacterial↗

[An epidemiological survey of group A, B, C & G hemolytic streptococci isolated from the Throat of Diabetic Children Streptococcicosis Study Group].

The incidence of group A, B, C & G hemolytic streptococci residing in the throat of children with diabetes was surveyed. The survey was carried out in August, 1982 and August, 1983. Included in the surveys were 136 diabetic children. The survey was also carried out with healthy school children at the same term as the contrast. The detection rate of the streptococci among the diabetic children was significantly higher than the healthy school children. A great portion of streptococci detected from these diabetic children was classified into group B, on the other hand, from the healthy school children, group A. Although from healthy school children, BIII, BIa and several other types were isolated, from diabetic children only BIa and BIII were isolated. The detection rate of the streptococci among the diabetic children was significantly higher in the early days in camp for improvement of diabetes than in the last day. In the latter case the detection rate was similar to that among healthy children. In the other words following antidiabetic therapy as diabetes improved the detection rate were also improved. Among these 136 diabetic children examined 31 gave positive cultures at both surveys each year. Serogroups or serotypes in each year examinations, isolates, were identical in 26 cases. Most of these groups were group B and the types were Ia and III.

Diabetes Mellitus↗

[Rapid diagnosis of influenza infection by PCR method--detection of influenza virus HA gene in throat swab].

We studied the detection of the HA gene of human influenza viruses in throat swabs obtained from the outbreaks of influenza in school children utilizing the polymerase chain reaction (PCR) method. Sensitivity and specificity of the PCR method was compared to conventional virus isolation using MDCK cells. Three pairs of primers for PCR in detecting the HA genes of AH1, AH3, and B influenza viruses showed both subtype and type specificity. The dilution experiments showed that influenza viruses, as few as 1.1-3.5 plaque-forming units per 50 microliters, were sufficient for the detection of HA genes by PCR method and the detection rate by PCR method was 2-3 fold higher than that by conventional method. Our results showed that the PCR method was a fast, sensitive and reliable method for the diagnosis of influenza infections.

Base Sequence↗

[Mycoplasma pneumoniae detection from throat swab by two-step polymerase chain reaction].

Two-step polymerase chain reaction (PCR) with primers designated against 16S rRNA gene of Mycoplasma pneumoniae for diagnosis of infection was evaluated in comparison with the conventional single-step PCR and culture methods. The two-step PCR method showed specific amplification of M. pneumoniae DNA and higher sensitivity (1.5 fg/assay) than the single-step PCR method. With the two-step PCR method, 76 of 322 throat swabs (23.6%) from patients with acute respiratory complaints gave positive results whereas 20.2% were positive in the culture method. Seven of 13 samples which were negative in the single-step PCR method but positive in either serological or the culture method showed positive results by the two-step PCR method. In addition, 5 samples which were weakly positive in the single-step PCR method showed distinctly positive results in the two-step PCR. These results indicate that the two-step PCR method is a useful tool for detection of M. pneumoniae in clinical specimens, although it requires a relatively sophisticated in technique.

Humans↗

Virulence of Yersinia pseudotuberculosis isolated from pork and from the throats of swine.

Yersinia pseudotuberculosis was isolated from retail pork and from healthy swine throats. These wild-type strains and their representative cured isogenic strains were tested for the presence of plasmids and several virulence factors, and these characteristics were compared with those of virulent strains from humans. Two pork isolates (serotype IVB) and four swine isolates (serotypes IIB, IIC, III, and IVB) harbored a 42- to 48-megadalton plasmid which had similar fragmentation patterns resulting from digestion with restriction endonuclease. These six strains were lethal for mice via oral challenge and were positive in autoagglutination and calcium dependency tests. They also invaded HeLa cells and induced cytotoxicity. Histopathological examination and indirect fluorescent-antibody staining provided definite evidence of the pathogenicity of these strains when tissue sections from orally infected mice were used. The virulence factors of wild-type pork and swine isolates with the 42- to 48-megadalton plasmid were identical to those of two human isolates (serotypes IVB and VB). Hence, these pork and swine isolates should be considered potentially pathogenic for humans. The finding suggests that retail pork and swine may play an important role in the epidemiology of human infections caused by Y. pseudotuberculosis.

Agglutination Tests↗

Significance of hemolytic colonies in throat cultures.

These studies indicate that a single strain of hemolytic streptococci almost exclusively predominates the bacterial flora in patients with streptococcal infections and in the carrier state. One can proceed with confidence that, in isolating streptococci from throat swabs cultured on blood-agar plates, only a single hemolytic colony need be picked for serological grouping and typing.

Carrier State↗

Serogrouping single colonies of beta-hemolytic streptococci from primary throat culture plates with nitrous acid extraction and Phadebact streptococcal reagents.

The serogrouping of isolated beta-hemolytic streptococcal colonies from throat cultures by a micronitrous acid extraction method employing Phadebact Streptotoccus Test reagents was compared with results obtained with the direct-plate Phadebact procedure and the autoclave and Streptomyces albus enzyme-lysozyme extraction methods. These data were compared with those from the Lancefield grouping obtained with a capillary precipitin test. The micronitrous acid extraction method was modified to yield an uncomplicated method that provides specific coagglutination responses from one beta-hemolytic streptococcal colony that may be collected from a primary blood agar plate either on the end of an applicator stick or from a sweep of an inoculating loop from mixed growth.

Agglutination↗

Evaluation of techniques for isolation of group A streptococci from throat cultures.

In the first study, selective sulfamethoxazole-trimethoprim blood agar (SXT-BA) and conventional blood agar (BA) plates incubated under CO2 and anaerobically were compared for their ability to recover group A streptococci from throat cultures. Recovery rates were: SXT-BA (anaerobic), 100%; SXT-BA (CO2), 98%; BA (anaerobic), 89.2%; and BA (CO2), 76.5%. Primary plate bacitracin test results could be read on significantly more of the SXT-BA plates. Readability rates were: SXT-BA (anaerobic), 97%; SXT-BA (CO2), 96%; BA (anaerobic), 70.6%; and BA (CO2), 32.4%. A second study compared with the SXT-BA method versus a BA-double-disk (BA-DD) method which utilizes conventional media with addition of a bacitracin differentiation and a sulfamethoxazole-trimethoprim susceptibility disk placed adjacent to one another in the heavy area of inoculation. Isolation rates were: SXT-BA, 100% and BA-DD, 88%. Readability rates for direct bacitracin tests were: SXT-BA, 92% and BA-DD, 76.2%. In our hands, the SXT-BA method was superior for yielding highest isolation rates and for yielding highest readability rates of direct bacitracin test results.

Adolescent↗

Rapid biochemical tests for the identification of groups A, B, C, F, and G streptococci from throat cultures.

A test employing three fluorogenic 4-methylumbelliferyl substrates and the lectin of Dolichos biflorus was developed for the identification of beta-hemolytic streptococcal colonies associated with throat cultures. This non-serological method is unique in that it permits the accurate identification of groups C, F, and G streptococci, as well as groups A and B streptococci. The method is rapid, simple, and specific and appears to be a useful means to identify groups A, B, C, F, and G streptococci.

Acetylglucosaminidase↗

Effect of delay in processing on the performance of Directigen for the detection of group A streptococci in throat swabs.

We compared a latex agglutination method, Directigen (Hynson, Westcott & Dunning) with a culture-based system for detection of group A streptococci in 964 throat swabs. Discordants were resolved by Lancefield typing of beta-hemolytic colonies. After culture, swabs were randomly assigned to be tested by Directigen within 4 h of sampling or after overnight storage. For swabs that yielded more than 50 beta-hemolytic colonies from culture, there was a highly significant difference (P less than 0.01) in sensitivity between the group tested early and the group tested after storage overnight. For swabs tested within 4 h of sampling, a trend (P = 0.07) existed between sensitivity and length of delay before testing.

Humans↗

Human Fc(gamma) receptors for differentiation in throat cultures of group C "Streptococcus equisimilis" and group C "Streptococcus milleri".

The biochemical characteristics and the presence of human Fc(gamma) receptors of 52 throat isolates of group C beta-hemolytic streptococci were examined. Among these isolates, 38 were identified as "Streptococcus milleri" and 14 were identified as "Streptococcus equisimilis." The differentiation of group C "S. equisimilis" from "S. milleri" with identical group antigens was easy to perform by the measurement of the size of the hemolytic zone on a sheep blood agar plate in an anaerobic atmosphere and by biochemical tests (Voges-Proskauer test). A clear-cut criterion for differentiation was noted among these isolates, i.e., the presence of Fc(gamma) receptors. "S. equisimilis," which are generally associated with pharyngitis, possess human Fc(gamma) receptors, while "S. milleri", which are generally isolated from healthy persons, have no such receptors.

Humans↗

Latex agglutination testing directly from throat swabs for rapid detection of beta-hemolytic streptococci from Lancefield serogroup C.

A latex agglutination method for the rapid detection of beta-hemolytic streptococci from Lancefield serogroup C in throat swabs from 403 university students with symptomatic pharyngitis was evaluated. Compared with culture, the rapid test was poorly sensitive (34.4%) but very specific (98.4%) in detecting group C beta-hemolytic streptococci. The sensitivity of the rapid test improved with an increasing quantity of growth on culture.

Adolescent↗

Comparison of three methods for detection of group A streptococci in throat swabs.

Group A streptococci are generally detected in throat swabs by (i) rapid antigen tests, (ii) conventional culture, or (iii) combinations of both. Direct fluorescent-antibody testing of a 2-h enrichment broth (FA/EN) was an accepted method for same-day results before the advent of rapid antigen tests. We compared FA/EN in Todd-Hewitt Broth (THB) with conventional culture and a rapid antigen test, TestPack Strep A (TPS). Nine hundred seventy specimens were evaluated in this study. Cultures were performed for 48 h on sheep blood agar (SBA) incubated aerobically and on a selective agar for group A streptococci (SSA) incubated in 5 to 10% CO2. Following a 2-h incubation, the fluorescent-antibody test was performed. A subculture of the centrifuged sediment from the THB enrichment was also done. In comparison with a positive culture on SBA or SSA or subculture of the THB pellet, the sensitivities and specificities of the different methods were as follows: SBA, 92 and 100%; SSA, 92 and 100%; TPS, 68 and 99%; FA/EN, 88 and 98%. The FA/EN method offers the potential for definitive finalized reports on the same day as specimen collection with greater sensitivity than TPS. This study included sequential plating and rapid antigen testing of a single swab. In a separate set of experiments to validate this study design, it was shown that recovery of streptococci from swabs plated sequentially on five plates did not vary with the order of plating and the actual proportion of organisms recovered from a swab on a single plate was only 1%.

Antigens, Bacterial↗

Prevalence of Taiwan variant of Epstein-Barr virus in throat washings from patients with head and neck tumors in Taiwan.

The prevalence of the Epstein-Barr virus (EBV) Taiwan variant was investigated in the throat washing (TW) samples from patients with head and neck tumors, persons with nonmalignant diseases, and healthy adults in Taiwan. By using the EBV (BNLF-1 gene)-specific primers and PCR, the EBV latent membrane protein gene BNLF-1 was detected in 91 (61%) of the 150 TW samples from patients with tumors, including 25 (78%) of 32 patients with nasopharyngeal carcinoma and 66 (56%) of 118 other patients with head and neck tumors. The TW samples from the 26 patients with nonmalignant tumors and 53 healthy adults were also examined. Approximately 47% of these samples were positive for the EBV gene. The PCR products of the BNLF-1 gene were then subjected to XhoI digestion. Sixty-eight of 91 PCR products (75%) showed the loss of the XhoI site, which indicated the presence of a Taiwan strain of EBV in patients with tumors. The DNA sequence of the BNLF-1 gene of the Taiwan variant revealed that the loss of the XhoI site was due to a nucleotide change from a G to a T at position 169,426 in comparison with the sequence of prototype EBV B95-8 cells. Furthermore, the Taiwan strain appeared significantly more frequently in the TWs and tissue samples from patients with nasopharyngeal carcinoma (88%; P < 0.001) and laryngeal carcinoma (80%; P < 0.02) than in those samples from healthy adults (about 40%). These data indicate that a Taiwan variant of EBV may be closely associated with head and neck tumors and suggest that this variant may be important in the pathogenesis of head and neck tumors.

Base Sequence↗

Evaluation of two rapid antigen assays, BioStar Strep A OIA and Pacific Biotech CARDS O.S., and culture for detection of group A streptococci in throat swabs.

Two rapid methods, BioStar Strep A OIA (OIA; BioStar, Inc., Boulder, Colo.), an optical immunoassay, and CARDS O.S. (O.S.; Pacific Biotech, Inc., San Diego, Calif.), a color immunochromographic assay, and two culture methods, one with 5% sheep blood agar (SBA) and one with Todd-Hewitt broth (TH; Remel, Lenexa, Kans.), were evaluated for use in the detection of Streptococcus pyogenes from pharnygeal swabs. Seven hundred forty-six double swabs (Culturette II) were processed, with OIA and SBA culture performed on one swab and O.S. and SBA culture performed on the other swab. The pledget from the Culturette II was incubated overnight in TH and was subcultured onto SBA for an additional 48 h in ambient air. All beta-hemolytic streptococci from culture were tested by a direct fluorescent-antibody test (Difco Laboratories, Detroit, Mich.). Specimens with discordant fluorescent-antibody test and rapid test results were also tested by using the Streptex latex agglutination reagent (Murex Diagnostics Limited, Dartford, England). The results obtained by all testing methods were compared with a combined test result ("gold standard"), which was defined as any positive culture detected by the SBA or TH culture methods and confirmed by Streptex latex agglutination or, in the case of negative results by both culture methods, a concomitant positive result by OIA and O.S. antigen testing. Sensitivity and specificity results for each of the methods were as follows, respectively: OIA, 81.0 and 97.5%; O.S., 74.4 and 99.0%; SBA culture, 92.3 and 98.3%; and TH culture, 86.4 and 100%. Both OIA and O.S. are suitable screening methods for detecting S. pyogenes directly from throat swabs but are of insufficient sensitivity to eliminate the need for backup cultures for specimens with negative OIA and O.S. results.

Antigens, Bacterial↗

Comparison of three methods for culturing throat swabs from cystic fibrosis patients.

In patients with cystic fibrosis who do not produce sputum, deep throat swabs are cultured for potential respiratory pathogens. Usually these swabs are directly streaked onto selective agar media. In a study of 50 pediatric cystic fibrosis patients, we compared this traditional method using rayon swabs with two methods having quantitative modifications: calcium alginate swabs eluted in Ringer's lactate and rayon swabs eluted in normal saline. The eluates were then processed quantitatively (three-step dilution series). The yield of potential pathogens was significantly higher with the two quantitative methods. Overall, the combination of alginate with Ringer's lactate was superior to the combination of rayon with saline, although only some of these differences achieved statistical significance.

Adolescent↗