Evaluation of the NucliSens basic kit for detection of Chlamydia trcachomatis and Neisseria gonorrhoeae in genital tract specimens using nucleic acid sequence-based amplification of 16S rRNA
Evaluation of the NucliSens basic kit for detection of Chlamydia trcachomatis and Neisseria gonorrhoeae in genital tract specimens using nucleic acid sequence-based amplification of 16S rRNA
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DOI：
10.1128/jcm.39.4.1429-1435.2001
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发表时间：
2001-04-01
影响因子：
9.4
通讯作者：
Kapala, J
中科院分区：
文献类型：
--
作者：
Mahony, JB;Song, X;Kapala, J
我们评估了一种新的RNA扩增和检测试剂盒，NucliSens基本试剂盒（Organon Teknika），用于检测泌尿生殖道标本中的沙眼衣原体和淋病奈瑟菌。基本试剂盒为RNA扩增和检测提供了一个开放平台，包含用于核酸提取的分离试剂、基于核酸序列的扩增（NASBA）试剂（酶和缓冲液）以及用于扩增产物电致发光（ECL）检测的通用荧光素标记探针。使用新鲜纯化和滴定的C. trachomatis和N.淋病和体外产生的RNA转录物进行敏感性测定时，基本试剂盒检测到1个包涵体形成单位的C。沙眼衣原体、N.淋病，和100个16 S rRNA的RNA分子的两种细菌。通过对250份样本进行N.用培养和NASBA法对96例淋病奈瑟氏菌进行了检测。用PCR和NASBA法检测沙眼衣原体，基本试剂盒检出142株沙眼衣原体中的139株。淋球菌培养阳性标本中检出100例，培养阴性标本中检出73例，敏感性为97.9%，特异性为98.7%。梭对于沙眼衣原体，基本试剂盒检出24/24份PCR阳性标本，71/72份PCR阴性标本，敏感性和特异性分别为100%和98.6%。基本试剂盒还检测到含有N.淋球菌和C.沙眼衣原体，使用多重NASBA测定，使用两种细菌的引物。NucliSens Basic Kit为开发敏感的性传播疾病RNA检测试剂盒提供了一个多功能平台。
We evaluated a new RNA amplification and detection kit, the NucliSens Basic Kit (Organon Teknika), for the detection of Chlamydia trachomatis and Neisseria gonorrhoeae in genitourinary specimens. The Basic Kit provides an open platform for RNA amplification and detection and contains isolation reagents for nucleic acid extraction, nucleic acid sequence-based amplification (NASBA) reagents (enzymes and buffers), and a generic ruthenium-labeled probe for electrochemiluminescent (ECL) detection of amplified product. Using freshly purified and titrated stocks of C. trachomatis and N. gonorrhoeae and in vitro-generated RNA transcripts for sensitivity determinations, the Basic Kit detected 1 inclusion-forming unit of C. trachomatis, I CFU of N. gonorrhoeae, and 100 RNA molecules of 16S rRNA for both bacteria. The clinical performance of the Basic Kit was evaluated by testing a total of 250 specimens for N. gonorrhoeae by culture and NASBA and a total of 96 specimens for C. trachomatis by PCR and NASBA, The Basic Kit detected 139 of 142 N. gonorrhoeae culture-positive specimens and gave a negative result for 73 of 74 culture-negative specimens, for a sensitivity and specificity of 97.9 and 98.7%, respectively. For C. trachomatis, the Basic Kit detected 24 of 24 PCR-positive specimens and gave a negative result for 71 of 72 PCR-negative specimens, for a sensitivity and specificity of 100 and 98.6%, respectively. The Basic Kit also detected specimens containing both N. gonorrhoeae and C. trachomatis, using a multiplex NASBA assay using primers for both bacteria. The NucliSens Basic Kit offers a versatile platform for the development of sensitive RNA detection assays for sexually transmitted diseases.