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[两种用于临床实践中分枝杆菌鉴定的反向杂交方法的比较]

[Comparison of two reverse hybridization methods for mycobacterial identification in clinical practice].

作者信息

Trombert-Paolantoni S, Poveda J-D, Figarella P

机构信息

Laboratoire Pasteur-Cerba biology, rue de l'Equerre, ZI des béthunes, 95066 Cergy-Pontoise, France.

出版信息

Pathol Biol (Paris). 2004 Oct;52(8):462-8. doi: 10.1016/j.patbio.2004.07.018.

Abstract

AIM OF THE STUDY

To evaluate the performance of two commercial methods for identification of Mycobacterium species: InnoLiPA Mycobacteria first version (Innogenetics) versus Genotype MTBC and Genotype Mycobacterium (HAIN) on, respectively, 2123 and 2164 distinct isolates.

MATERIALS AND METHODS

Both techniques are based on the reverse hybridization of PCR products to their complementary probes immobilized on membrane strips. The InnoLiPA assay targets the 16S-23S rRNA spacer region. The HAIN test is composed of two kits: Genotype MTBC, for identification of tuberculosis complex mycobacteria, is based on gyrB DNA sequence polymorphism. Genotype Mycobacterium kit targets the 23S rDNA for identification of mycobacteria other than tuberculosis (MOTT) and tuberculosis complex mycobacteria. Both assays identify complex tuberculosis mycobacteria and respectively, eight and 12 species of MOTT. Moreover, the Genotype MTBC allows species differentiation within the M. tuberculosis complex.

RESULTS

Eighty-eight percent and 95% of mycobacteria were identified by InnoLiPA and HAIN, respectively. Hybridization remained negative for 11% of isolates with InnoLiPA and 4% with HAIN. An identification of MOTT was obtained by conventional identification in all cases after the use of InnoLiPA. MOTT and one M. tuberculosis was obtained after HAIN procedure. Unidentified species were complementary to a specific probe in 5% of the cases with InnoLiPA and 17% with HAIN.

CONCLUSION

HAIN identifies more mycobacteria species than does InnoLiPA and allows identification in the M. tuberculosis complex. However, failure in identification occurs only with MOTT with InnoLiPA when one M. tuberculosis was found among mycobacteria non identified with HAIN.

摘要

研究目的

评估两种用于鉴定分枝杆菌菌种的商业方法的性能:InnoLiPA分枝杆菌第一代版本(Innogenetics公司)与Genotype MTBC和Genotype Mycobacterium(HAIN公司),分别针对2123株和2164株不同的分离株。

材料与方法

两种技术均基于PCR产物与固定在膜条上的互补探针的反向杂交。InnoLiPA检测法靶向16S - 23S rRNA间隔区。HAIN检测法由两个试剂盒组成:Genotype MTBC用于鉴定结核分枝杆菌复合群,基于gyrB DNA序列多态性。Genotype Mycobacterium试剂盒靶向23S rDNA用于鉴定非结核分枝杆菌(MOTT)和结核分枝杆菌复合群。两种检测法均可鉴定结核分枝杆菌复合群以及分别8种和12种MOTT。此外,Genotype MTBC可区分结核分枝杆菌复合群内的菌种。

结果

InnoLiPA和HAIN分别鉴定出88%和95%的分枝杆菌。InnoLiPA检测中11%的分离株杂交结果为阴性,HAIN检测中为4%。使用InnoLiPA后,所有病例经传统鉴定均获得MOTT鉴定结果。HAIN检测后获得MOTT和1株结核分枝杆菌鉴定结果。InnoLiPA检测中5%的病例以及HAIN检测中17%的病例,未鉴定出的菌种与特定探针互补。

结论

HAIN比InnoLiPA能鉴定出更多的分枝杆菌菌种,且能在结核分枝杆菌复合群中进行鉴定。然而,当在HAIN未鉴定出的分枝杆菌中发现1株结核分枝杆菌时,InnoLiPA仅在MOTT鉴定中出现失败情况。

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