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RNA amplification by nucleic acid sequence-based amplification with an internal standard enables reliable detection of Chlamydia trachomatis in cervical scrapings and urine samples.
J Clin Microbiol. 1996 Dec; 34(12):3108-14.JC

Abstract

In the present study, the suitability of RNA amplification by nucleic acid sequence-based amplification (NASBA) for the detection of Chlamydia trachomatis infection was investigated. When comparing different primer sets for their sensitivities in NASBA, use of both the plasmid and omp1 targets resulted in a detection limit of 1 inclusion-forming unit (IFU), while the 16S rRNA appeared to be the most sensitive RNA target for amplification (10(-3) IFU). In contrast, for DNA amplification by PCR, the plasmid target was optimal (10(-2) IFU), which is 10 times less sensitive than rRNA NASBA. To exclude false negativity in NASBA detection because of inhibition of amplification and/or inefficient sample preparation, an internal standard was developed. The internal control was added prior to sample preparation. This 16S rRNA NASBA with an internal control was compared with a plasmid DNA PCR by using a group of C. trachomatis-negative (n = 41) and -positive (n = 37) cervical scrapings, as determined by enzyme immunoassay (EIA). In addition, urine samples from the EIA-positive women were tested (n = 17). Both NASBA and PCR assays were able to detect C. trachomatis in all EIA-positive cervical scrapings, the corresponding urine samples, and two samples from the EIA-negative group. The internal NASBA standard was found clearly in all EIA-negative samples. In conclusion, these results indicate that detection of C. trachomatis by RNA amplification by NASBA with an internal standard is a suitable and highly sensitive detection method, with potential use in the diagnosis of urogenital C. trachomatis infections with cervical scrapings as well as urine specimens.

Authors+Show Affiliations

Department of Pathology, University Hospital Vrije Universiteit, Amsterdam, The Netherlands.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Comparative Study
Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

8940456

Citation

Morré, S A., et al. "RNA Amplification By Nucleic Acid Sequence-based Amplification With an Internal Standard Enables Reliable Detection of Chlamydia Trachomatis in Cervical Scrapings and Urine Samples." Journal of Clinical Microbiology, vol. 34, no. 12, 1996, pp. 3108-14.
Morré SA, Sillekens P, Jacobs MV, et al. RNA amplification by nucleic acid sequence-based amplification with an internal standard enables reliable detection of Chlamydia trachomatis in cervical scrapings and urine samples. J Clin Microbiol. 1996;34(12):3108-14.
Morré, S. A., Sillekens, P., Jacobs, M. V., van Aarle, P., de Blok, S., van Gemen, B., Walboomers, J. M., Meijer, C. J., & van den Brule, A. J. (1996). RNA amplification by nucleic acid sequence-based amplification with an internal standard enables reliable detection of Chlamydia trachomatis in cervical scrapings and urine samples. Journal of Clinical Microbiology, 34(12), 3108-14.
Morré SA, et al. RNA Amplification By Nucleic Acid Sequence-based Amplification With an Internal Standard Enables Reliable Detection of Chlamydia Trachomatis in Cervical Scrapings and Urine Samples. J Clin Microbiol. 1996;34(12):3108-14. PubMed PMID: 8940456.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - RNA amplification by nucleic acid sequence-based amplification with an internal standard enables reliable detection of Chlamydia trachomatis in cervical scrapings and urine samples. AU - Morré,S A, AU - Sillekens,P, AU - Jacobs,M V, AU - van Aarle,P, AU - de Blok,S, AU - van Gemen,B, AU - Walboomers,J M, AU - Meijer,C J, AU - van den Brule,A J, PY - 1996/12/1/pubmed PY - 1996/12/1/medline PY - 1996/12/1/entrez SP - 3108 EP - 14 JF - Journal of clinical microbiology JO - J. Clin. Microbiol. VL - 34 IS - 12 N2 - In the present study, the suitability of RNA amplification by nucleic acid sequence-based amplification (NASBA) for the detection of Chlamydia trachomatis infection was investigated. When comparing different primer sets for their sensitivities in NASBA, use of both the plasmid and omp1 targets resulted in a detection limit of 1 inclusion-forming unit (IFU), while the 16S rRNA appeared to be the most sensitive RNA target for amplification (10(-3) IFU). In contrast, for DNA amplification by PCR, the plasmid target was optimal (10(-2) IFU), which is 10 times less sensitive than rRNA NASBA. To exclude false negativity in NASBA detection because of inhibition of amplification and/or inefficient sample preparation, an internal standard was developed. The internal control was added prior to sample preparation. This 16S rRNA NASBA with an internal control was compared with a plasmid DNA PCR by using a group of C. trachomatis-negative (n = 41) and -positive (n = 37) cervical scrapings, as determined by enzyme immunoassay (EIA). In addition, urine samples from the EIA-positive women were tested (n = 17). Both NASBA and PCR assays were able to detect C. trachomatis in all EIA-positive cervical scrapings, the corresponding urine samples, and two samples from the EIA-negative group. The internal NASBA standard was found clearly in all EIA-negative samples. In conclusion, these results indicate that detection of C. trachomatis by RNA amplification by NASBA with an internal standard is a suitable and highly sensitive detection method, with potential use in the diagnosis of urogenital C. trachomatis infections with cervical scrapings as well as urine specimens. SN - 0095-1137 UR - https://www.unboundmedicine.com/medline/citation/8940456/RNA_amplification_by_nucleic_acid_sequence_based_amplification_with_an_internal_standard_enables_reliable_detection_of_Chlamydia_trachomatis_in_cervical_scrapings_and_urine_samples_ L2 - http://jcm.asm.org/cgi/pmidlookup?view=long&pmid=8940456 DB - PRIME DP - Unbound Medicine ER -