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
S A MorréA J van den Brule

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, a...Continue Reading

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Citations

Oct 16, 2015·São Paulo Medical Journal = Revista Paulista De Medicina·Carolina Sanitá Tafner FerreiraMárcia Guimarães da Silva
Jan 20, 2006·Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America·Matthew J BurtonRobin L Bailey
Aug 23, 2005·Emerging Infectious Diseases·Servaas A MorréA Salvador Peña
Nov 5, 2005·Applied and Environmental Microbiology·Khaled H Abd el-GalilAshok Mulchandani
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Nov 26, 2016·International Journal of STD & AIDS·Christian T BautistaJose L Sanchez
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Oct 17, 2018·Sensors·Shrikant Dashrath WarkadTaisun Kim
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