Abstract
One hundred fifty-three blood samples from patients positive for the human immunodeficiency virus (HIV) were analyzed by polymerase chain reaction (PCR) to detect the presence ofMycobacterium avium. Samples were collected from patients who also had blood cultures performed by a radiometric method. Blood samples were centrifuged on a Ficoll-Hypaque gradient to purify peripheral blood mononuclear cells. The purified cells were washed and incubated with a resin, boiled to release mycobacterial DNA, and then amplified. Polymerase chain reaction products were detected by a nonisotopic method. A 123 base-pair (bp) insertion sequence, namely IS6110, fromMycobacterium tuberculosis complex was also included in the reaction as an internal control ofTaq polymerase activity to exclude the presence of enzyme inhibitors. This IS6110 fragment can be distinguished from the 383 bp target product on ethidium bromide-stained agarose gel and may also be used in a colorimetric assay. Such results were compared with the results of culture and indicated that the assay is as sensitive as bacteriological methods, though faster.
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Kiehn TE, Edwards FF, Brannon P Tsang AY, Maio M, Gold JWM, Whimbey E, Wong B, McClatchy JK, Armstrong DA: Infections caused byMycobacterium avium complex in immunocompromised patients: diagnosis by blood culture and fecal examination, antimicrobial susceptibility tests, and morphological and seroagglutination characteristics. Journal of Clinical Microbiology 1985, 21: 168–173.
Nightingale SD, Byrd LT, Southern PM, Jockusch JD, Cal SX, Wynne BA: Incidence ofMycobacterium avium-intracellulare complex bacteremia in human immunodeficiency virus-positive patients. Journal of Infectious Diseases 1992, 165: 1082–1085.
Wallace JM, Hannah J:Mycobacterium avium complex in patients with the acquired immunodeficiency synrome. Chest 1988, 93: 926–932.
Young LS, Inderlied CB, Berlin OG, Gottlieb MS: Mycobacterial infections in AIDS patients, with emphasis on theMycobacterium avium complex. Reviews of Infectious Diseases 1986, 8: 1024–1033.
Horsbourgh CR:Mycobacterium avium complex infec tion in the acquired immunodeficiency syndrome. New England Journal of Medicine 1991, 324: 1332–1338.
Nussbaum JM, Dealist C, Lewis W, Heseltim PNR: Rapid diagnosis by buffy coat smear of disseminatedMycobacterium avium complex infection in patients with acquired immunodeficiency syndrome. Journal of Clinical Microbiology 1990, 28: 631–632.
Gill VJ, Park CH, Stock F, Gosey LL, Witebsky FG, Masur H: Use of lysis-centrifugation (Isolator) and radiometric (BACTEC) blood culture systems for the detection of mycobacteremia. Journal of Clinical Microbiology 1985, 22: 543–546.
Colombrita D, Ravizzola G, Pinsi G, Li Vigni R, Pirali F, Turano A: Rapid detection and identification of mycobacteria from blood of patients with acquired immune deficiency syndrome. Journal of Medical Microbiology 1990, 32: 271–273.
Patel RJ, Fries JWU, Piessens WF, Wirth DF: Sequence analysis and amplification by polymerase chain reaction of a cloned DNA fragment for identification ofMycobacterium tuberculosis. Journal of Clinical Microbiology 1990, 28: 513–518.
Portillo PD, Murillo LA, Patarroyo ME: Amplification of a species-specific DNA fragment ofMycobacterium tuberculosis and its possible use in diagnosis. Journal of Clinical Microbiology 1991, 29: 2163–2168.
Sritharan V, Barker RH Jr: A simple method for diagnosingM. tuberculosis infection in clinical samples using PCR. Molecular Cellular Probes 1991, 5: 385–395.
Brisson-Noel A, Aznar C, Chureau C, Nguyen S, Pierre C, Bartoli M, Bonete R, Pialoux G, Gicquel B, Garrigue G: Diagnosis of tuberculosis by DNA amplification in clinical practice evaluation. Lancet 1991, 338: 364–366.
Kwok S, Higuchi R: Avoiding false positives with PCR. Nature 1989, 339: 237–238.
Singer-Sam J, Tanguay RL, Riggs AD: Use of Chelex to improve the PCR signal from a small number of cells. Amplifications 1989, 3: 11.
Walsh PS, Metzeger DA, Higuchi R: Chelex-100TM as a medium for simple extraction of DNA for PCR-based typing from forensic material. BioTechniques 1991, 10: 506–513.
Hance AJ, Grandchamp B, Levy-Frebault V, Lecassier D, Rauzier J, Bocart D, Gicquel B: Identification of mycobacteria by amplification of mycobacterial DNA. Molecular Microbiology 1989, 3: 843–849.
Eisenach KD, Cave MD, Bates JH, Crowford JT: Polymerase chain reaction amplification of a repetitive DNA sequence specific forMycobacterium tuberculosis. Journal of Infectious Diseases 1990, 161: 977–981.
Thierry D, Cave MD, Eisenach KD, Crowford JT, Bates JM, Gicquel B, Guesdon JL: IS6110, an IS-like element of theMycobacterium tuberculosis complex. Nucleic Acids Research 1990, 18: 188.
Mantero G, Zonaro A, Bertolo P, Primi D: DNA enzyme immunoassay (DEIA): a general method for detecting polymerase chain reaction products based on anti-DNA antibody. Clinical Chemistry 1991, 37: 422–429.
Masur H, and the Public Health Task Force on Prophylaxis and Therapy forMycobacterium avium complex: Special report. Recommendations on prophylaxis and therapy for disseminatedMycobacterium avium complex disease in patients infected with the human immunodeficiency virus. New England Journal of Medicine 1993, 329: 898–905.
Hoy J, Mijch A, Sandland M, Grayson L, Lucas R, Dwyer B: Quadruple-drug therapy forMycobacterium aviumintracellulare bacteremia in AIDS. Journal of Infectious Diseases 1990, 161: 801–805.
Noordhoek GT, Kolk AHJ, Bjune G, Catty D, Dale JW, Fine PEM, Godfrey-Faussett P, Cho SN, Shinnick T, Svenson SB, Wilson S, Van Embden JDA: Sensitivity and specificity of PCR for detection ofMycobacterium tuber culosis: a blind comparison study among seven laboratories. Journal of Clinical Microbiology 1994, 32: 277–284.
Eisenach KD, Cave MD, Bates JR, Crowford JT: Detection ofMycobacterium tuberculosis in sputum samples using a polymerase chain reaction. American Reviews of Respiratory Diseases 1991, 144: 1160–1169.
De Wit D, Wootton M, Allan B, Steyn L: Simple method for production of internal control DNA forMycobacterium tuberculosis polymerase chain reaction assays. Journal of Clinical Microbiology 1993, 31: 2204–2207.
Kolk AHJ, Noordhoek GT, De Leeuw O, Kuijper S, van Embden JDA:Mycobacterium smegmatis strain for detection ofMycobacterium tuberculosis by PCR used as internal control for inhibition of amplification and for quantification of bacteria. Journal of Clinical Microbiology 1994, 32: 1354–1356.
Fries JWU, Patel RJ, Piessens WF, Wirth DF: Genus and species-specific DNA probes to identify mycobacteria using the polymerase chain reaction. Molecular Cellular Probe 1990, 4: 87–105.
Fries JWU, Patel RJ, Piessens WF, Wirth DF: Detection of untreated mycobacteria by using polymerase chain reaction and specific probes. Journal of Clinical Microbiology 1991, 29: 1744–1747.
Boddinghaus B, Rogall T, Flohr T, Blocker H, Bottger ER: Detection and identification of mycobacteria by amplification of rRNA. Journal of Clinical Microbiology 1990, 28: 1751–1759.
Kulski JK, Khinsoe C, Pryce T, Christiansen K: Use of a multiplex PCR to detect and identifyMycobacterium avium andM. intracellulare in blood culture fluids of AIDS patients. Journal of Clinical Microbiology 1995, 33: 668–674.
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De Francesco, M.A., Colombrita, D., Pinsi, G. et al. Detection and identification ofMycobacterium avium in the blood of AIDS patients by the polymerase chain reaction. Eur. J. Clin. Microbiol. Infect. Dis. 15, 551–555 (1996). https://doi.org/10.1007/BF01709362
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DOI: https://doi.org/10.1007/BF01709362