I. Objectives Self-etching primer adhesive system is affected to dentin surface conditioning and priming. Especially application time of self-etching primer is very important factor of clinical procedure which has direct influence on smear layer, etching reaction and primer penetration to dentin. This study evaluated the influence of application time of self-etching primers on microtensile bond strength (${\mu}{\;}TBS$) to dentin using three self-etching primer adhesive systems.(omitted)
Backgrounds : Recent technological developments have introduced a new method to identifying M. tuberculosis complex DNA in clinical samples directly. The direct amplification test (DAT) is approved for identifying M. tuberculosis complex in respiratory specimens that are smear-positive for acid-fast bacilli (AFB). When there is a discrepancy between the AFB smear and DAT, no information on their clinical utility is currently available. In this study, the diagnostic reliability of DAT was investigated in suspected pulmonary tuberculosis patients whose sputum AFB smear was negative. Methods : From June 1, 1998 through May 30, 1999, 909 patients with presumed active pulmonary tuberculosis were enrolled. A sputum AFB stain, culture, DAT and/or biopsy were performed. Using the criteria of clinical tuberculosis or confirmed tuberculosis, the positive predictive value of DAT in diagnosing pulmonary tuberculosis was investigated. Results : The positive predictive value of DAT was 82.1% by the clinically active tuberculosis criteria. However, it decreased to 61.5% when diagnosis was restricted to only to culture positive or biopsy proven cases. The false positive rate of DAT was 18.0%. Conclusion : The DAT is a valuable diagnostic method in suspected patients whose sputum AFB is was negative.
Background: Diagnosis by direct microscopy and/or by culture of the Mycobacterium tuberculosis from body fluids or biopsy specimens is "Gold standard". However, the sensitivity of direct microscopy after Ziehl-Neelsen staining is relatively low and culture of mycobacteria is time consuming. Detection of mycobacterial DNA in clinical samples by the polymerase chain reaction is highly sensitive but laborious and expensive. Therefore, rapid, sensitive and readily applicable new tests need to be developed. So we had evaluated the clinical significance of serologic detection of antibody to 38 kDa antigen, which is known as the most specific to the M. tuberculosis complex, and culture filtrate antigen by ELISA in sputum AFB smear negative patients. Method: In this study, culture tests for acid fast bacilli with sputa or bronchial washing fluids of 183 consecutive patients who were negative of sputum AFB smear were performed. Simultaneously serum antibodies to 38 kDa antigen and unheated culture filtrate of M. tuberculosis were detected by an ELISA method. Results: The optical densities of ELISA test with 38 kDa and culture filtrate antigen were significantly higher in active pulmonary tuberculosis cases than in non tuberculous pulmonary diseases (p<0.05), but in patients with active pulmonary tuberculosis, those of the sputum culture positive patients for M. tuberculosis were not significantly different from those of the sputum culture negative cases(p>0.05). In the smear-negative active pulmonary tuberculosis patients, the sensitivity of the ELISA using 38 kDa antigen and culture filtrate was 20.0% and 31.4%. respectively. The specificity was 95.3% and 93.9%. respectively. Conclusion : In active pulmonary tuberculosis but smear negative, the serologic detection of antibody to 38 kDa antigen and culture filtrate by ELISA cannot substitute traditional diagnostic tests and does not have clinically significant role to differenciate the patient with active pulmonary tuberculosis from other with non-tuberculous pulmonary diseases.
Background : Rifampicin(RFP) is a key component of the antituberculous short-course chemotherapy and the RFP resistance is a marker of multi-drug resistant(MDR) tuberculosis. RPoB gene encodes the $\beta$-subunit of RNA polymerase of M. tuberculosis which is the target of RFP. And rpoB gene mutations are the cause of RFP resistance of M. tuberculosis. Although several reports showed that PCR-SSCP would be a rapid diagnostic method for identifying the RFP resistance, there were few reports Performed using direct, clinical specimens. So we Performed PCR-SSCP analysis of rpoB gene of M. tuberculosis in direct, clinical specimens. Methods : 75 clinical specimens were collected from patients at Asan Medical Center from June to August 1996. After PCR of IS 6110 fragments, 43 both AFB smear-positive and IS6110 fragment PCR-positive specimens were evaluated. The RFP susceptibility test was referred to the referral laboratory of the Korean Tuberculosis Institute. DNA was extracted by bead beater method. And heminested PCR was done using 0.1ul(1uCi) [$\alpha-^{32}P$]-dCTP. SSCP analysis was done using non-denaturating MDE gel electrophoresis. Results : The results of PCR of IS6110 fragments of M. tuberculosis were positive in 55(73%) cases of 75 AFB smear-positive clinical specimens. Of the 55 specimens, RFP susceptibility was confirmed in only 43 specimens. Of the 43 AFB smear-positive and IS6110 fragment-positive specimens, 29 were RFP susceptible and 14 were RFP resistant. All the RFP susceptible 29 strains showed the same mobility compared with that of RFP sensitive H37Rv in SSCP analysis of ropB gene. And all the other RFP resistant 13 strains showed the different mobility. In other words they showed 100% identical results between PCR-SSCP analysis and traditional susceptibility test. Conclusion : The PCR-sseP analysis of rpoB gene in direct clinical specimens could be used as a rapid diagnostic method for detecting RFP resistant M. tuberculosis.
Perazzi, Beatriz E.;Menghi, Claudia I.;Coppolillo, Enrique F.;Gatta, Claudia;Eliseth, Martha Cora;De Torres, Ramon A.;Vay, Carlos A.;Famiglietti, Angela M. R.
Parasites, Hosts and Diseases
/
v.48
no.1
/
pp.61-65
/
2010
The objectives of this study were to conduct a prevalence survey of trichomoniasis in pregnant women and to evaluate the utility of different methods for its diagnosis. A total of 597 vaginal exudates from pregnant women who were examined at the Hospital de Clinicas in Buenos Aires, Argentina from 1 August 2005 to 31 January 2007, were prospectively and consecutively evaluated. The investigation of Trichomonas vaginalis was made by different microscopic examinations, and culture on liquid medium. The sensitivity and specificity of the microscopic examinations were assessed considering culture on liquid medium as the "gold standard". The prevalence of T. vaginalis obtained by culture on liquid medium was 4.0% (24/597). The prevalence of T. vaginalis obtained by direct wet smear, prolonged May-Grunwald Giemsa staining, and sodium acetate-formalin (SAF)/methylene blue staining-fixing technique was 1.8%, 2.3% and 2.5%, respectively. The sensitivity of the direct wet smear was 45.8%, that of the prolonged May-Grunwald Giemsa staining was 58.3%, and that of the SAF/methylene blue method was 62.5%. Considering the 3 microscopic examinations altogether, the sensitivity rose to 66.7% and the specificity was 100% for all of them. This is the first time that the prevalence data of T. vaginalis by culture in pregnant women are published in Argentina. Due to the low sensitivity obtained by microscopy in asymptomatic pregnant women, the use of the liquid medium is recommended during pregnancy, in order to provide an early diagnosis and treatment.
Five cases infected with Rhabditis sp. were detected in a survey to examine the stool specimens from rural primary school children. A large number of the larvae of Rhabditis sp. detected by the direct cellophane thick smear were cultured by the alter paper method. The eBamination was carried out in April through June, 1980 in Tangjeong-Myon, Ahsan.Gun, and in September 1983 in Sandong-Eup, Yeongwol-Gun. The results obtained in this study were as follows: 1. Prevalence: Out of 925 children, 5(0.54%) children were found to be positive. The number of the detected larvae was 110/0.1gm of feces in case 1, 35 in case 2, 130 in case 3, 86 in case 4 and 62 in case 5. 2. Larvae: The larvae were prepared by means of the direct fecal smear and measured by a micrometer equipped in the microscope. Twelve (12) through 15-day old larvae in culture were $197.1{\mu}{\textrm{m}}$ long in average, and the maximum size of the matured stage larvae was $884.0{\times}25.9{\mu}{\textrm{m}}$. However, the length variation was ranged as 173.0 to $884.0{\mu}{\textrm{m}}$. 3. Adults: The size of clubbed adult female was $1, 357{\mu}{\textrm{m}}(1, 176~1, 419)$ in length and $80{\mu}{\textrm{m}}(79~82)$ in width. Length of buccal cavity was $33{\mu}{\textrm{m}}$. A long cylindrical esophagus($273{\mu}{\textrm{m}}$) of the worms with a valved posterior cardiac bulb and with median bulbar swelling was morphologically indicated. Ditance from mouth to vulva was occupied 58% of body length. Male worm was $1, 006{\mu}{\textrm{m}}(890~1, 148)$ in length and $49{\mu}{\textrm{m}}(48~49)$ wide. Caudal alee of bursa and spicules ($75{\mu}{\textrm{m}}$ in length) were well developed. 4. Eggs: The oval shaped eggs in the female uterus, when cultured, were $66{\times}56{\mu}{\textrm{m}}$ in sixte, and the eggs laid by the adult could not be detected. So, reproduction might be thought to be ovoviviparity. 5. The five cases were re-examined during the period from the 1st to the 3rd week after stool examinations, but Rhabditis sp. were detected again in 4 cases in 1st week. When they were examined in 3rd week, larvae could not be detected. So, it was thought that the infection of Rhabditis sp. to humans was facultative.
The use of screening tests as part of a diagnostic work-up is common in domestic canine practice, but understanding of the diagnostic test characteristics and factors affecting diagnostic accuracy is not clear among clinicians. This article was aimed to provide clinicians with a better understanding on the selection of test kits and with a proper interpretation of test results using an example from heartworm(Dirofilaria immitis) studies. From the literatures, diagnostic accuracy varied depending on the kits: percent average sensitivity and specificity of ELISA antigen-detecting kits were DiroChek(Synbiotics, USA) 78.1 and 95.2, SNAP(IDEXX, USA) 66.3 and 98.1, and Solo Step(Heska, Switzerland) 69.5 and 97.5, respectively, while the values for three hematological methods(Modified Knott's, direct smear and capillary tube) ranged from 38.4 to 81.8% and from 96.9 to 100%, respectively. Furthermore, it was also reported that the prevalence of heartworm disease in domestic dog populations varied depending on the regions studied: 2.5-22.8% for microfilarial test and 2.2-66.3% by ELISA. The values of predictive values for positive(PPV) and negative(NPV) provide useful information to clinicians on the probability of heartworm infection, but the PPV and NPV are greatly dependent on the heartworm prevalence. This suggests that PPV or NPV values of a test should be interpreted carefully in different clinical settings. Practical methods on the interpretation taking into account heartworm prevalence were discussed.
Chong, Yun-Sop;Kim, Yoon-Chung;Kim, Byung-Soo;Yi, Kui-Nyung;Lee, Sam-Uel Y.
The Journal of the Korean Society for Microbiology
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v.13
no.1
/
pp.1-5
/
1978
Listeria monocytogenes infection was considered a rather rare disease and occurs mostly either in newborn babies or in young children. However, there has been increasing reports of this infection in elderly person with various underlying disease. Recently we have experienced two cases of Listeria meningitis; one in a 4-year-old male with an acute lymphoblastic leukemia, and the other in a 43-year-old female with a breast cancer. Both were on various chemotherapeutic agents for their primary diseases when the organism, L. monocytogenes was found in their celebospinal fluid(CSF). The degree of CSF pleocytosis were quite different by cases. The former case showed a marked increase, $3,350/mm^3$, and the latter slight, $410/mm^3$, Both showed a slight decrease of CSF glucose ranging 39 to 43mg/100ml. It seems that a routine CSF analysis bears a limitted value in the diagnosis or Listeria meningitis. A direct smear of CSF with Gram's stain revealed gram-positive bacilli in one case, but none in the other. Bacterial culture of CSF yielded plenty colonies in one case, but a few in the other. It seems that isolation of L. monocytogenes must not be considered very easy, and a negative direct smear does not necessarily mean a negative culture. The two isolates we obtained showed the typical cultural and biochemical characteristics of L. monocytogenes and were found to belong to serotypes 1b and 4b. It was our experience that the identification of this organism was not very much matter because of its distinct characteristics, but the most important matter was how to think of the possibility of this organism at the begining. The two isolates were both susceptible to cephalothin, chloramphenicol, erythromycin, tetracycline and gentamicin; intermediate to ampicillin, penicillin and kanamycin; and resistant to cloxacillin.
In dogs, correct diagnosis of estrus is important and the exact time of ovulation can be determined by variouse methods. Vaginal cytology has commonly used in conjunction with the physical examination, clinical history, vaginoscopy, and hormonal assays to determine the stage of the reproductive cycle. This study was therefore investigated the effectiveness of direct ovulation detector designed by changes of electrical resistance in vaginal mucus following different estrus cycles with several methods; vaginal cytology, concentration of plasma estrogen and progesterone, and direct examination by laparotomy. A total of 12 bitches was selected for the study and observed estrus signs. The bitches were evaluated clinical sign (vulvar swelling and bleeding), cytological examination (keratocyte and RBC), electrical resistance, plasma estrogen and progesterone concentration for estrus assessment. Accuracy of ovulation detection by vaginal cytology was significantly (p<0.05) lower than those by electrical resistance and plasma progesterone concentration, based on the confirmation by laparotomy. Vaginal smear is not confidential method compared to detection of electrical resistance and plasma progesterone concentration at ovulation. Although the value of electrical resistance was varied at the same points of estrus in individuals, ovulation was occurred at the first day which shown the peak of electrical resistance and mating time was third day after peak. In conclusion, ovulation detector designed by changes of electrical resistance is an effective and economic instrument for predicting estrus and ovulation in bitches.
To evaluate the efficacy of ronidazole for treatment of Tritrichomonas foetus infection, 6 Tritrichomonas-free kittens were experimentally infected with a Korean isolate of T. foetus. The experimental infection was confirmed by direct microscopy, culture, and single-tube nested PCR, and all cats demonstrated trophozoites of T. foetus by day 20 post-infection in the feces. From day 30 after the experimentally induced infection, 3 cats were treated with ronidazole (50 mg/kg twice a day for 14 days) and 3 other cats received placebo. Feces from each cat were tested for the presence of T. foetus by direct smear and culture of rectal swab samples using modified Diamond's medium once a week for 4 weeks. To confirm the culture results, the presence of T. foetus rRNA gene was determined by single-tube nested PCR assay. All 3 cats in the treatment group receiving ronidazole showed negative results for T. foetus infection during 2 weeks of treatment and 4 weeks follow-up by all detection methods used in this study. In contrast, rectal swab samples from cats in the control group were positive for T. foetus continuously throughout the study. The present study indicates that ronidazole is also effective to treat cats infected experimentally with a Korean isolate of T. foetus at a dose of 50 mg/kg twice a day for 14 days.
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