• Title/Summary/Keyword: Negative staining

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A Comparision of the Sensitivities of Culture, Chlamydiazyme and Direct Immunofluorescence Staining for Chlamydia trachomatis in Non-gonococcal Urethritis (비임균성 뇨도염 환자에서 Chlamydia trachomatis 검출방법에 관한 연구 (배양법, 효소면역법 및 직접면역형광법의 비교 관찰))

  • Choi, Tae-Yeal;Kim, Choon-Won;Kim, Jung-Hwan
    • The Journal of the Korean Society for Microbiology
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    • v.21 no.3
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    • pp.393-397
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    • 1986
  • Chlamydia trachomatis is one of the most common cause in non-gonococcal urethritis. There are several diagnostic methods for Chlamydia trachomatis; culture method using McCoy cell, enzyme immunoassay and direct immunofluorescence staining etc. We have studied a sensitivities of culture, chlamydiazyme and direct immunofluorescence staining(DIF). 85 male patients previously conformed to non-gonococcal urethritis have been selected in this study. Three samples were concurrently collected in the same patient. First sample was used to inoculation in McCoy cell, 2'nd sample was used to Chlamydiazyme test and 3'rd sample was used to direct immunofluorescence staining method. The results are following. 1. All culture, Chlamydiazyme and DIF positive cases are 15/85(17.7%). 2. Culture and Chlamydiazyme positive but DIF negative cases absent. 3. Culture and DIF positive, but Chlamydiazyme negative cases are 2/85(2.4%). 4. Chlamydiazyme and DIF positive, but culture negative cases are 9/85(10.6%). 5. Culture positive, but Chlamydiazyme and DIF negative cases are 6/85(7.1%). 6. Chlamydiazyme positive, but culture and DIF negative cases are 7/85(8.2%). 7. DIF positive, but culture and Chlamydiazyme negative cases are 3/85(3.5%). 8. All culture, Chlamydiazyme and DIF negative cases are 43/85(50.1%). In summarized, anyone positive cases of culture, Chlamydiazymc and DIF are 42/85(49.9%).

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Detection of Mycobacterium tuberculosis by PCR from Trace Clinical Specimens and Paraffin-embedded Tissues (임상가검물과 파라핀 포매 조직에서 PCR법을 이용한 결핵균의 검출)

  • 김은중;최우순;황석연
    • Biomedical Science Letters
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    • v.6 no.1
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    • pp.55-63
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    • 2000
  • This study has been carried out to investigate the sensitivity of polymerase chain reaction (PCR) method over conventional acid-fast bacilli (AFB) staining and/culture methods for the detection of Mycobacterium tuberculosis from trace body fluid and paraffin-embedded tissues (PET) specimens. A total of 65 cases were employed for the AFB staining and culture test, and a total of 50 cases were subjected to PCR and histopathological analysis. Among the specimen showing negative reaction to AFB staining, 12.1% were positive to PCR and 3.7% of the specimen representing negative result to culture test showed positive reaction to PCR. In addition, 20.0% of the specimen with AFB negative showed positive reaction to PCR. From these results, it could be concluded that PCR method overwhelms AFB staining and culture tests in sensitivity and specificity to M tuberculosis detection.

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An Electron Microscopic Structure of Rotavirus by Negative Stain (Negative stain을 이용한 Rotavirus의 투과전자현미경적 구조)

  • Kwon, Jung-Kyun
    • Applied Microscopy
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    • v.24 no.4
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    • pp.107-114
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    • 1994
  • The Rotaviruses are members of the family Reoviridae and are the major cause of severe childhood gastroenteritis worldwide. Recently, electron microscopy has been used to detect non-group A rotaviruses to determine a relatively high resolution structure of the rotavirion. Mature, infectious virions(double-shelled particles) have a diameter of approximately 70nm, and have a capsid structure composed of two concentric protein layers. We have studied patient's stool specimen by negative staining technique complete removal of sucrose suspension. This negative staining technique that could be carried out in about 30 minutes and that could be used with crude stool specimen was an advantage of major significance. Removal of sucrose in the sample by has been completed washing with distilled of sucrose and by washing with distilled water. Ultrastructurally, typical feature of rotavirus has a double capsid construction with an inner capsid of 55nm and on outer 65-70nm diameter can be clearly demonstrated.

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Lack of Prognostic Significance of C-erbB-2 Expression in Low- and High- grade Astrocytomas

  • Muallaoglu, Sadik;Besen, Ali Ayberk;Ata, Alper;Mertsoylu, Huseyin;Arican, Ali;Kayaselcuk, Fazilet;Ozyilkan, Ozgur
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.3
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    • pp.1333-1337
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    • 2014
  • Background: Astrocytic tumors, the most common primary glial tumors of the central nervous system, are classified from low to high grade according to the degree of anaplasia and presence of necrosis. Despite advances in therapeutic management of high grade astrocytic tumors, prognosis remains poor. In the present study, the frequency and prognostic significance of c-erb-B2 in astrocytic tumors was investigated. Materials and Methods: Records of 72 patients with low- and high-grade astrocytic tumors were evaluated. The expression of C-erbB-2 was determined immunohistochemically and intensity was recorded as 0 to 3+. Tumors with weak staining (1+) or no staining (0) were considered Her-2 negative, while tumors with moderate (2+) and strong (3+) staining were considered Her-2 positive. Results: Of the 72 patients, 41 (56.9%) had glioblastoma (GBM), 10 (13.9%) had diffuse astrocytoma, 15 (20.8%) had anaplastic astrocytoma, 6 (8.3%) had pilocytic astrocytoma. C-erbB-2 overexpression was detected in the tumor specimens of 17 patients (23.6%). Six (8.3%) tumors, all GBMs, exhibited strong staining, 2 (2.7%) specimens, both GBMs, exhibited moderate staining, and 9 specimens, 5 of them GBMs (12.5%), exhibited weak staining. No staining was observed in diffuse astrocytoma and pilocytic astrocytoma specimens. Median overall survival of patients with C-erbB-2 negative and C-erbB-2 positive tumors were 30 months (95%CI: 22.5-37.4 months) and 16.9 months (95%CI: 4.3-29.5 months), respectively (p=0.244). Conclusions: Although there was no difference in survival, C-erbB-2 overexpression was observed only in the GBM subtype.

Different Profiles of the Negatively Stained Citrus Canker Bacterium Xanthomonas citri pv. citri Depending on Culture Media and Heavy Metal Stains

  • Kim, Ki-Woo;Lee, In-Jung;Hyun, Jae-Wook;Lee, Yong-Hoon;Park, Eun-Woo
    • The Plant Pathology Journal
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    • v.26 no.1
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    • pp.90-92
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    • 2010
  • Staining profiles and bacterial morphology were compared in Xanthomonas citri pv. citri by a transmission electron microscopy. Four types of negative staining regimes were employed depending on culture media and heavy metal stains. The bacterial cells grown on LB agar media often appeared clustered on the supporting film. Meanwhile, individual bacterial cells could be readily found on the preparations from LB broth media. Typical rod-shaped cells (ca. $1\;{\mu}m$ in length) and their flagella were observed in either 2% uranyl acetate (UA) or 2% neutralized potassium phosphotungstate (PTA) staining. The UA-stained bacteria often showed relatively intact cell morphology and rather positively stained cells with a thin electron-dense stain depth around bacteria. The PTA-stained bacteria were characterized by the wrinkled cell surface where the stain was entrapped in grooves. In addition, distinct electron-dense stain depth was evident around the PTA-stained preparations. Numerous fimbriae could be mostly observed from the PTA-stained preparations of the two culture media, but not from the UA-stained preparations.

Comparison of Pre-Stain Suspension Liquids in the Contrasting Ability of Neutralized Potassium Phosphotungstate for Negative Staining of Bacteria

  • Kim, Ki-Wooh;Jung, Woo-Kyung;Park, Yong-Ho
    • Journal of Microbiology and Biotechnology
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    • v.18 no.11
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    • pp.1762-1767
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    • 2008
  • Image contrast of whole bacteria was compared in Staphylococcus aureus and Escherichia coli depending on pre-stain suspension liquids by energy-filtering transmission electron microscopy. The two bacterial strains were suspended in three most commonly used liquids for negative staining (triple distilled water [DW], phosphate-buffered saline [PBS], and nutrient broth [NB]) and directly observed without staining or stained with neutralized potassium phosphotungstate (PTA), respectively. Even though in low contrast, unstained bacteria were observed owing to their inherent electron density and cell shape in zero-loss (elastic scattering) images. After being suspended in PBS, unstained bacteria appeared to have higher contrast and more refined periphery than DW-suspended ones, and extracellular appendage structures such as fimbriae and flagella could be discerned. The unstained bacteria appeared to be invariably surrounded with electron-lucent precipitates, possibly from PBS. As far as delineation of the structures, the combination of DW or PBS suspension with subsequent staining provided the most satisfactory results, as evidenced by the high contrast of bacterial morphology and appendage structures. However, after being suspended in NB and stained with PTA, bacteria often had too high contrast or poor staining, with electron-dense aggregates around the bacteria. These results suggest that suspension with concentrated organic aliquots including broth media before PTA staining could deteriorate image contrast, and should be used only in dilute form for visualizing bacterial morphology and appendage structures. Moreover the contrast enhancement of unstained bacteria by salt granules would be advantageous in demonstrating bacterial sorption of environmental particles like heavy metals, maintaining minimal contrast for cell imaging.

Application of alkaline phosphatase staining of cytology specimen for differential diagnosis of canine osteosarcoma (세포 검사시료에서 alkaline phosphatase 염색법을 활용한 개 골육종의 감별 진단)

  • Park, Byoung-Yong;Park, Chul;Cho, Ho-Seong
    • Korean Journal of Veterinary Service
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    • v.34 no.4
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    • pp.397-401
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    • 2011
  • Aspiration of lytic bone lesions is an excellent diagnostic test in the initial evaluation of primary bone tumor. However, cytologically, it can be difficult to differentiate osteosarcoma (OSA) from other bone neoplasms, including fibrosarcoma, chondrosarcoma, synovial cell sarcoma, malignant fibrous histiocytoma and malignant peripheral nerve sheath tumor. The purpose of this study is to introduce alkaline phosphatase (ALP) staining to differentiate OSA from other mesenchymal tumors. Tumors actively producing bone are specifically positive for ALP staining. Unstained, cytologic specimens were incubated for 10 minutes with nitroblue tetrazolium chloride/5-bromo-4-chloro-3-indolyl phosphate toluidine salt-phosphatase substrate. Among 20 cases of cytology specimen, 14 were positive for ALP staining and histopathology, 6 were negative for ALP staining and histopathology. ALP staining was 100% sensitive and specificity for the diagnosis of OSA. Aspirate cytology with ALP staining was a simple, fast, safe and accurate diagnostic test for the evaluation of suspected OSA lesions in dogs.

Metachronous extranodal natural killer/T-cell lymphoma of nasal type and primary testicular lymphoma

  • Maeng, Young-In;Lee, Sun-Jae
    • Journal of Yeungnam Medical Science
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    • v.38 no.3
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    • pp.231-234
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    • 2021
  • We report a rare case of metachronous lymphoma with two distinct cell lineages in a 75-year-old man. The patient complained about having nasal obstruction for 2 years and extranodal natural killer (NK)/T-cell lymphoma of the nasal type was diagnosed from a biopsy. The immunohistochemical staining for CD56 and in situ hybridization for Epstein-Barr virus (EBV)-encoded small RNA (EBER-ISH) were positive and the tumor cells were negative for CD20. After 13 months of concurrent chemoradiotherapy, the patient presented with swelling of the left testis. Positron emission tomography scan detected an abnormal uptake in the testis. A diffuse large B-cell lymphoma, not otherwise specified, was diagnosed from subsequent radical orchiectomy. The immunohistochemical staining revealed to be positive for CD20, BCL2, BCL6, and MYC and negative for CD10 and EBER-ISH.

Detection of viral-like particles in feces of Korean indigenous calves with diarrhea by negative staining (음성대조 염색법을 이용한 설사 한우송아지 분변내 바이러스양 입자 검색)

  • 임종수;강춘원;이태욱;김내영;정용운;강문일;한동운;최현성;이채용
    • Korean Journal of Veterinary Service
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    • v.22 no.1
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    • pp.43-52
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    • 1999
  • Presently, viral isolation in the diarrheal feces can be reached by many tools such as fluorescent antibody test(FA), negative contrast electron microscopy(NCEM), virus neutralization test, cell culture, and so on. The purpose of the study was to aimed at the establishment of simplified NCEM technique which can be efficiently applied for diarrheal feces and also the understanding on prevalence of viral-induced diarrhea in calves. One hundred fourty-seven korean indigenous calves with diarrhea were examined to their feces by the modified NCEM. Among them, 98(66.7%) were confirmed to have one or more viruses in feces. The viruses detected were identified as rotavirus(33.3%), coronavirus(16.3% ), togavirus(10.2%) and herpesvirus(0.7%). Ten cases of combined viral infection were consisted of 8 with rotavirus+coronavirus, one with rotavirus+togavlrus and one with rotavirus+herpesvirus. Dirrheal types could classified by yello-wish watery(44.9a ), blood-tinged(19.7% ), white watery(17.7% ) , brownish watery(14.3%), greenish watery(3.4%) diarrhea, respectively. Yellowish watery diarrhea(66cases) was frequently included rotavirus(31.8%), coronavirus(15.2%), and togavirus(13.6%), respectively. Consequently, these results suggest that the modified NCEM is reliable and efficient diagnostic tool for detection of viruses in the diarrheal feces and many calves rearing in Chonnam province have been exposed to some enteric viral agents mainly including rotavirus and coronavirus.

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Cytomegalovirus-associated esophageal ulcer in an immunocompetent infant: When should ganciclovir be administered?

  • Jang, Hyo-Jeong;Kim, Ae Suk;Hwang, Jin-Bok
    • Clinical and Experimental Pediatrics
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    • v.55 no.12
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    • pp.491-493
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    • 2012
  • Cytomegalovirus (CMV)-associated esophageal ulcer is rare in immunocompetent infants. The presence of inclusion bodies and immunohistochemical staining for CMV in biopsy specimens obtained during esophagogastroduodenoscopy (EGD) indicate that such ulcers occur because of CMV infection. A 7-week-old female infant who experienced frequent vomiting and feeding intolerance was diagnosed with a massive CMV-associated ulcer in the distal esophagus. The ulcer improved after conservative treatment using proton-pump inhibitors; however, ganciclovir was not administered. In a follow-up EGD biopsy specimen, no CMV inclusion bodies were present, and immunohistochemical staining results for this virus were negative. The presence of CMV inclusion bodies indicates active viral replication. If persistent inclusion bodies or positive immunohistochemical staining for CMV is observed in follow-up biopsy specimens, ganciclovir may be used to treat CMV-associated esophageal ulcers.