{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,12,14]],"date-time":"2025-12-14T10:45:05Z","timestamp":1765709105814,"version":"3.48.0"},"reference-count":15,"publisher":"BMJ","issue":"9","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["J Clin Pathol"],"published-print":{"date-parts":[[1996,9]]},"abstract":"<jats:p>AIMS: To investigate the effectiveness of digoxigenin incorporated double stranded DNA probes produced by the polymerase chain reaction (PCR), for the detection of Pneumocystis carinii, using in situ hybridisation (ISH). METHODS: Formalin fixed, paraffin wax embedded sections of 26 human lung samples from 11 patients with P carinii pneumonia (PCP), and 15 with various types of fungal and viral pneumonia, were obtained during necropsy or transbronchial lung biopsy. Three additional PCP induced rat lung samples were also tested. PCR probes were prepared using the digoxigenin labelling mixture, and they were amplified from the DNA of a PCP induced rat lung after administration of dexamethasone, on the basis that 5S ribosomal RNA sequences are identical in human and rat P carinii. ISH was performed using this probe, and visualised using the digoxigenin nucleic acid detection kit. An immunohistochemical study using anti-human Pneumocystis monoclonal antibody was also carried out in parallel. RESULTS: ISH positively stained eight (of eight) lung necropsy specimens from patients with PCP, three (of three) transbronchial lung biopsy specimens from patients with PCP, and none of the three PCP induced rat lung specimens. In contrast, none of the specimens from patients with pneumonia caused by Aspergillus sp (n = 5), Candida sp (n = 4), Cryptococcus sp (n = 2), mucormycosis (n = 2), or cytomegalovirus (n = 2) were positive on ISH or immunohistochemistry. CONCLUSIONS: Using a digoxigenin labelled PCR probe for the entire 5S rRNA sequence in conjunction with conventional staining, ISH is highly reactive and specific for the diagnosis of PCP.<\/jats:p>","DOI":"10.1136\/jcp.49.9.712","type":"journal-article","created":{"date-parts":[[2007,6,9]],"date-time":"2007-06-09T00:50:46Z","timestamp":1181350246000},"page":"712-716","source":"Crossref","is-referenced-by-count":25,"title":["Simple detection of the 5S ribosomal RNA of Pneumocystis carinii using in situ hybridisation."],"prefix":"10.1136","volume":"49","author":[{"given":"M","family":"Kobayashi","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"T","family":"Urata","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"T","family":"Ikezoe","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"E","family":"Hakoda","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Y","family":"Uemura","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"H","family":"Sonobe","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Y","family":"Ohtsuki","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"T","family":"Manabe","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"S","family":"Miyagi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"I","family":"Miyoshi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"239","published-online":{"date-parts":[[1996,9,1]]},"reference":[{"key":"ref_1","first-page":"65","article-title":"Pneumocystis carinii","author":"Hopkin, J.M.","year":"1991","unstructured":"Hopkin JM. Pneumocystis carinii. Oxford: Oxford University Press, 1991:65-6."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"589","DOI":"10.1056\/NEJM198803103181001","article-title":"Diagnosis of Pneumocystis carinii pneumonia: improved detection in sputum with use of monoclonal antibodies","volume":"318","author":"Kovacs, J.A.; Ng, V.L.; Masur, H.; Leoung, G.; Hadley, W.K.; Evans, G.","year":"1988","unstructured":"Kovacs JA, Ng VL, Masur H, Leoung G, Hadley WK, Evans G, et al. Diagnosis of Pneumocystis carinii pneumonia: improved detection in sputum with use of monoclonal antibodies. N EnglJ Med 1988;318:589-93.","journal-title":"N EnglJ Med"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1016\/0022-1759(87)90435-2","article-title":"Detection of Pneumocystis carinii in lung derived samples using monoclonal antibody to a 82 kDa parasite antigen","volume":"98","author":"Linder, E.; Lundin, H.; Vorma, H.","year":"1987","unstructured":"Linder E, Lundin H, Vorma H. Detection of Pneumocystis carinii in lung derived samples using monoclonal antibody to a 82 kDa parasite antigen. J Immunol Methods 1987;98:57-62.","journal-title":"J Immunol Methods"},{"key":"ref_4","first-page":"387","article-title":"Immunohistochemical detection of Pneumocystis carinii in transbronchial lung biopsy specimens: Antigen difference between human and rat Pneumocystis carinii","volume":"22","author":"Kobayashi, M.; Moriki, T.; Uemura, Y.; Takehara, N.; Kubonishi, I.; Taguchi, H.","year":"1992","unstructured":"Kobayashi M, Moriki T, Uemura Y, Takehara N, Kubonishi I, Taguchi H, et al. Immunohistochemical detection of Pneumocystis carinii in transbronchial lung biopsy specimens: Antigen difference between human and rat Pneumocystis carinii. Jpn J Clin Oncol 1992;22:387-92.","journal-title":"Jpn J Clin Oncol"},{"key":"ref_5","first-page":"37","article-title":"In vitro amplification and digoxigenin labeling of single stranded and double stranded DNA probes for diagnostic in situ hybridization","volume-title":"PCR: clinical diagnostics and research","author":"Finckh, U.; Lingenfelter, P.A.; Henne, K.W.; Schmidt, C.A.; Siegert, W.; Myerson, D.","year":"1992","unstructured":"Finckh U, Lingenfelter PA, Henne KW, Schmidt CA, Siegert W, Myerson D. In vitro amplification and digoxigenin labeling of single stranded and double stranded DNA probes for diagnostic in situ hybridization. In: Rolfs A, Schuller I, et al, eds. PCR: clinical diagnostics and research. Berlin: Springer Verlag, 1992:A37-43."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"859","DOI":"10.1093\/infdis\/158.4.859","article-title":"Cloning DNA from Pneumocystis carinii","volume":"158","author":"Wakefield, A.E.; Hopkin, J.M.; Burns, J.; Hipkiss, J.B.; Stewart, T.J.; Moxon, E.R.","year":"1988","unstructured":"Wakefield AE, Hopkin JM, Burns J, Hipkiss JB, Stewart TJ, Moxon ER. Cloning DNA from Pneumocystis carinii. J Infect Dis 1988;158:859-61.","journal-title":"J Infect Dis"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"519","DOI":"10.1038\/334519a0","article-title":"Ribosomal RNA sequence shows Pneumocystis carinii to be a member of fungi","volume":"334","author":"Edman, J.C.; Kavacs, J.A.; Masur, H.; Santi, D.V.; Elwood, H.J.; Sogin, M.L.","year":"1988","unstructured":"Edman JC, Kavacs JA, Masur H, Santi DV, Elwood HJ, Sogin ML. Ribosomal RNA sequence shows Pneumocystis carinii to be a member of fungi. Nature 1988; 334:519-22.","journal-title":"Nature"},{"key":"ref_8","first-page":"57680","article-title":"A novel diagnostic method of Pneumocystis carinii: In situ hybridization of ribosomal ribonucleic acid with biotinylated oligonucleotide probes","volume":"63","author":"Hayashi, Y.; Watanabe, J.; Nakata, K.; Fukayama, M.; Ikeda, H.","year":"1990","unstructured":"Hayashi Y, Watanabe J, Nakata K, Fukayama M, Ikeda H. A novel diagnostic method of Pneumocystis carinii: In situ hybridization of ribosomal ribonucleic acid with biotinylated oligonucleotide probes. Lab Invest 1990;63:57680.","journal-title":"Lab Invest"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1985","DOI":"10.1128\/JCM.29.9.1985-1990.1991","article-title":"Detection of Pneumocystis carinii sequences by polymerase chain reaction: animal models and clinical application to non-invasive specimens","volume":"29","author":"Kitada, K.; Oka, S.; Kimura, S.; Shimada, K.; Serikawa, T.; Yamada, J.","year":"1991","unstructured":"Kitada K, Oka S, Kimura S, Shimada K, Serikawa T, Yamada J, et al. Detection of Pneumocystis carinii sequences by polymerase chain reaction: animal models and clinical application to non-invasive specimens. Y Clin Microbiol 1991;29:1985-90.","journal-title":"Y Clin Microbiol"},{"key":"ref_10","first-page":"3","article-title":"In vitro cultivation","volume-title":"carinii pneumonia","author":"Armstrong, M.Y.K.; Cushion, M.T.","year":"1994","unstructured":"Armstrong MYK, Cushion MT. In vitro cultivation. In: Walzer PD, ed. Pneumocystis carinii pneumonia. 2nd edn. New York: Marcel Dekker Inc, 1994:3-24.","edition":"2nd edn"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1016\/0166-6851(89)90067-4","article-title":"Phylogenetic association of Pneumocystis carinii with the 'rhizopoda\/ myxomycota\/zygomycota group' indicated by comparison of 5S ribosomal RNA sequences","volume":"32","author":"Watanabe, J.; Hori, H.; Tanabe, K.; Nakamura, Y.","year":"1989","unstructured":"Watanabe J, Hori H, Tanabe K, Nakamura Y. Phylogenetic association of Pneumocystis carinii with the 'rhizopoda\/ myxomycota\/zygomycota group' indicated by comparison of 5S ribosomal RNA sequences. Mol Biochem Parasitol 1989;32:163-8.","journal-title":"Mol Biochem Parasitol"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"4881","DOI":"10.1093\/nar\/12.12.4881","article-title":"The nucleotide sequences of the 5S rRNAs of seven molds and a yeast and their use in studying ascomycete phylogeny","volume":"12","author":"Chen, M.-W.; Anne, J.; Volckert, G.; Huysmans, E.; Vandenberghe, A.; De Wachter, R.","year":"1984","unstructured":"Chen M-W, Anne J, Volckert G, Huysmans E, Vandenberghe A, De Wachter R. The nucleotide sequences of the 5S rRNAs of seven molds and a yeast and their use in studying ascomycete phylogeny. Nucleic Acids Res 1984; 12:4881-92.","journal-title":"Nucleic Acids Res"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1663","DOI":"10.1111\/j.1365-2958.1992.tb00891.x","article-title":"An Aspergillus fumigatus alkaline protease mutant constructed by gene disruption is defficient in extracellular activity","volume":"6","author":"Tang, C.M.; Cohen, J.; Holden, W.","year":"1992","unstructured":"Tang CM, Cohen J, Holden W An Aspergillus fumigatus alkaline protease mutant constructed by gene disruption is defficient in extracellular activity. Mol Microbiol 1992; 6:1663-71.","journal-title":"Mol Microbiol"},{"key":"ref_14","first-page":"61","article-title":"The theory and practice of in situ hybridization","volume-title":"situ hybridization: a practical approach","author":"Wilkinson, D.G.","year":"1992","unstructured":"Wilkinson DG. The theory and practice of in situ hybridization. In: Wilkinson DG, ed. In situ hybridization: a practical approach. Oxford: Oxford University Press, 1992:61-73."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"48","DOI":"10.1093\/ajcp\/103.1.48","article-title":"Rapid method for detection of Aspergillus 5S ribosomal RNA using a genus specific oligonucleotide probe","volume":"103","author":"Montone, K.T.; Litzky, L.A.","year":"1995","unstructured":"Montone KT, Litzky LA. Rapid method for detection of Aspergillus 5S ribosomal RNA using a genus specific oligonucleotide probe. Am Y Clin Pathol 1995;103:48-5 1.","journal-title":"Am Y Clin Pathol"}],"container-title":["Journal of Clinical Pathology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.1136\/jcp.49.9.712","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,12,14]],"date-time":"2025-12-14T10:43:29Z","timestamp":1765709009000},"score":1,"resource":{"primary":{"URL":"https:\/\/jcp.bmj.com\/lookup\/doi\/10.1136\/jcp.49.9.712"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[1996,9]]},"references-count":15,"journal-issue":{"issue":"9","published-online":{"date-parts":[[1996,9,1]]},"published-print":{"date-parts":[[1996,9]]}},"alternative-id":["10.1136\/jcp.49.9.712"],"URL":"https:\/\/doi.org\/10.1136\/jcp.49.9.712","relation":{},"ISSN":["0021-9746","1472-4146"],"issn-type":[{"type":"print","value":"0021-9746"},{"type":"electronic","value":"1472-4146"}],"subject":[],"published":{"date-parts":[[1996,9]]}}}