Chemiluminescence Detection Systems

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1. Dioxetanes

Alkaline phosphatase-dioxetane chemiluminescence systems have been used in a wide variety of DNA hybridization assays for detection of infectious


Selected Studies That Have Used Chemiiuminescence to Detect Viruses, Chlamydia trachomatis, and Other Microorganisms




Barley yellow dwarf virus

Bluetongue virus

Bovine enteric coronavirus

Bovine immunodeficiency-like virus

Bovine leukosis virus

Bursal disease virus

Chicken anemia virus


Dengue virus

Enterovirus (poliovirus)

Epstein-Barr virus

Feline infectious peritonitis virus Grapevine closterovirus Hepatitis B virus

Hepatitis C virus Herpes simplex virus

Human immunodeficiency virus










Jacobs et al. (1992)

Miliukiene et al. (1991)

Tham and Stanislawek (1992a,b) (DX)

Musiani et al. (1991a,1992) (DX); Yang et al. (1991) (DX)

Martinez and Weiss (1993) (AP)

Bronstein et al. (1989c) (DX); Farmar and Castañeda (1991) (DX); Yang et at. (1991) (DX)

Escarceller et al. (1992) (DX)

Khalil et al. (1991a,b) (AE); Bouveresse and Bourgeois (1992) (AP); Boxall (1992) (LU); McCartney et al. (1993) (LU)

Ireland and Samuel (1989) (LU); Robertson et al. (1991) (AE); Weare et al. (1991) (AE)

Geiger and Caselmann (1992) Khalil et al. (1991b) (AE)

Bronstein and Voyta (1989) (DX) Bronstein and Voyta (1989) (DX) Puchhammerstoeckl et al. (1993) (AP)

Pronovost et al. (1981) (ILU); Dalessio and Ashley (1992) (LU) Conway et al. (1990) (AP); Zachar et al. (1991) (DX)

Ou et al. (1990) (AE); Blackburn et al. (1991) (EL); Schmidt (1991) (AE); Schmidt and Gschnait (1991); Gudibande et al. (1992) (EL); Kenten et al. (1992) (EL); Suzuki et al. (1992) (DX); Rapier et al. (1993) (AE); Wages et al. (1993) (EL) Bettens et al. (1991) (DX) Ishii and Ghosh (1993) (AP)

Human papilloma virus

Human T-cell leukemia virus

Human T-cell lymphotropic virus Influenza virus

Lentivirus Parvovirus

Potato virus Y, potato spindle tuber viroid

Potato, pome fruit viroid

Respiratory syncytial virus, rotavirus

Rubella virus

Varicella zoster virus


Chlamydia trachomatis


Neisseria gonorrhoeae Plasmodium falciparum Toxoplasma gondii

IA Khalil et al. (1991b) (AE); Jacobs et al. (1992)

PCR/SH Balaguer et al. (1991b) (BL); Kenten et al. (1991) (EL)

ISH Hawkins and Cumming (1990) (LU)

MH McKimm-Breschkin (1992) (DX)

IM Arenkov et al. (1991)

MH Welnicki and Hiruki (1993) (AP)

MH Gustaferro and Persing (1992) (LU)

SH Clyne et al. (1989) (DX); Urdea et al. (1989) (DX); Gratton et al. (1990) (AE); Mercer et al. (1990) (AE); Iwen et al. (1991) (AE)

IA Neman-Simha et al. (1991); Dumornay et al. (1992) (AE); Jang et al. (1992) (AE); Scieux et al. (1992a,b) (AE)

SH Bull and Shanson (1992) (AE)

PCR/MH Sritharan and Barker (1991) (DX)

SH Urdea et al. (1989) (DX); Vlaspolder et al. (1993) (AE)

" Assay formats include: BH, bead-based hybridization; H, hybridization; IA, immunoassay; IM, immunity; ISH, in situ hybridization; MH, membrane-based hybridization; PCR, polymerase chain reaction; RT, reverse transcriptase; SD, strand displacement; SH, solution hybridization.

4 Chemiluminescent (CL) methods employed (if known): AE, acridinium ester; AP, alkaline phosphatase (most likely with dioxetane substrate); BL, bioluminescence; DX, dioxetane; EL, electrochemiluminescence; ILU, isoluminol; LU, enhanced luminol.

agents. Membrane-based hybridization assays have been used for the detection of HBV (Bronstein et al., 1989c; Yang et al., 1991; Escarceller et al., 1992), herpes simplex virus (HSV-1) (Bronstein and Voyta, 1989), CMV (Musiani et al., 1991a,1992; Yang et al., 1991), HIV-1 (Zachar et al., 1991), and other viral agents (Fouly et al., 1992; Tham and Stanislawek, 1992a,b; Fuchs et al., 1993). Solution hybridization assays include those for HBV (Urdea et al., 1990), HIV-1 (Suzuki et al., 1992), and Chlamydia (Clyne et al., 1989; Urdea et al., 1989). In situ hybridization assays have been performed with both HSV-1 infected cells (Bronstein and Voyta, 1989) and HIV-infected cells (Bronstein et al., 1989b). Finally, assays for retroviruses based on the detection of reverse transcriptase activity can be coupled with chemiluminescence detection by measuring the enzymatic incorporation of digoxigenin-labeled nucleotides with anti-digoxigenin alkaline phosphatase and a dioxetane substrate (Suzuki et al., 1993).

Commercially available detection systems incorporating dioxetanes include the AmpliProbe® system (ImClone Systems) for membrane-based hybridization assays for HBV, CMV, and EBV (Yang et al., 1991), the Hybrid Capture™ System HBV DNA Assay (Murex Diagnostics Ltd., Kent, UK), and solution hybridization assay systems for Chlamydia trachomatis and HBV detection (Chiron Corporation; Clyne et al., 1989; Urdea et al., 1989,1990).

2. Luminol

DNA hybridization assays using the ECL system with direct HRPlabeled probes include detection of bovine enteric Coronavirus in a slot blot hybridization assay (Collomb et al., 1992) and a solution-phase hybridization assay for HBV DNA (Urdea et al., 1987,1990). In situ hybridization for detection of human papillomavirus (HPV) type 16 has been performed with an indirect labeled probe (Hawkins and Cumming, 1990). ECL systems have also been used for the immunoassay detection of several viruses, including grapevine closterovirus (Pollini et al., 1993) and parvovirus B 19 (O'Neill and Coyle, 1992).

3. Acridinium Esters

DNA hybridization assays incorporating AE-labeled probes have been developed for detection of several infectious agents from clinical samples, including C. trachomatis, Neisseria gonorrhoeae, fungal pathogens, mycobacteria, and several common bacterial pathogens (Nelson and Kacian, 1990). These assay systems, called PACE 2™ and ACCUPROBE™, are available commercially through Gen-Probe, Inc. (San Diego, CA). The GenProbe system for screening for Chlamydia has been compared with both culture and nonculture antigen detection methods including enzyme immunoassays and immunofluorescent antibody tests (Gratton et al., 1990; Mercer et al., 1990; Iwen et al., 1991). The PACE 2™ system can provide a rapid, reliable alternative to culture and immunoassay methods for the detection of Chlamydia from cervical samples (Iwen et al., 1991). Solution hybridization (hybridization protection) assays with AE-labeled probes have been used for the detection of PCR-amplified HIV-1 DNA (Ou et al., 1990; Schmidt, 1991; Rapier et al., 1993).

In addition, AEs have also been used to label antibodies that have been incorporated into automated immunoassay formats for the detection of infectious agents and antibody screening from clinical samples (Khalil et al., 1991a,b).

4. Electrochemiluminescence

Electrochemiluminescence detection has been used in both manual (Blackburn et al., 1991; Gudibande et al., 1992; Kenten et al., 1992) and automated (QPCR System 5000; Wages et al., 1993) post-PCR amplification DNA hybridization assays for the detection of HIV-1 and HPV (Kenten et al., 1991).

5. Bioluminescence

Detection of asymmetric amplified papillomavirus sequences using solution-phase hybridization with a G6PDH-labeled oligonucleotide and solid-phase capture has been performed using a bioluminescence assay (Bala-guer et al., 1991b).

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