Typing of Neisseria gonorrhoeae Opa and NG-MAST gene of 12 pairs of sexual contact gonorrhea patients in China

Abstract

To identify the genomic species of Neisseria gonorrhoeae, evaluate the difference between two molecular epidemiological methods and examine the relationship between sex partners and genotypes of bacteria, 24 strains of Neisseria gonorrhoeae isolated from the outpatients with gonorrhea were identified by using the Opa genotyping and NG-MAST genotyping and the relationship between genotypes and phenotypes was studied. Twenty-four strains of Neisseria gonorrhoeae fell into 10 ST genotypes by NG-MAST genotyping, whereas these strains were classified into 12 OT Opa genotypes by Opa genotyping. A new epidemic strain of ST genotype (217-86% homologisation 178) in China was identified. It is concluded that genotypes of each pair of strains from a pair of patient/ sex partner besides 45/46 are the same, indicating that contagious infection take place between patient and the sex partner. Opa genotyping was more effective than NG-MAST genotyping in identifying the genomic species of Neisseria gonorrhoeae. ST genotype could be further classified into different Opa-types.

Review and International Recommendation of Methods for Typing Neisseria gonorrhoeae Isolates and Their Implications for Improved Knowledge of Gonococcal Epidemiology, Treatment, and Biology

“…Opa typing is based on the PCR amplification of 11 opa genes followed by restriction endonuclease digestion with TaqI and HhaI, separation of fragments by gel electrophoresis, and subsequent visualization of the banding patterns (118). The method imparts high typeability and reproducibility and excellent discriminatory ability, even in comparison with highly discriminatory DNA sequence-based approaches (19,20,26,49,65,71,74,93,96,104,118,121,127,174,182,186,189). However, Opa typing has the same disadvantages as RFLP methods, including labor-intensiveness, subjectiveness, and a need for pronounced standardization for interlaboratory comparisons.…”

“…However, Opa typing has the same disadvantages as RFLP methods, including labor-intensiveness, subjectiveness, and a need for pronounced standardization for interlaboratory comparisons. Opa typing has been used to define gonococcal populations within a geographic area, for the identification of clusters of strains (8,9,19,20,26,49,65,71,74,93,96,104,118,121,127,174,182,186,189), for tracing strain transmission between sexual contacts (9,19,74,186), for resolving suspicions of reinfection (102), for substantiation of treatment failure (77,102), and for detection of mixed infection (102). Opa typing has been used to increase the discriminatory power of other typing methods, including full-or extended-length porB sequencing or NG-MAST in specific situations, especially those involving extreme microepidemiological analysis (19,65,96,104,174,186) (Table 1).…”

“…In such cases, an analysis of the exact test and its costeffectiveness is warranted and should not be routine. Typing with different highly discriminatory genetic methods on the same N. gonorrhoeae strains generally displays similar levels of discrimination and relatively high congruence; however, there is rarely complete identity (9,19,24,49,51,54,58,65,69,71,73,82,83,96,104,115,121,125,135,153,172,174,176,178,180,182,184,186). This is not surprising since the different genetic targets of the typing methods follow divergent evolutionary pathways and consequently have different evolutionary histories.…”

Modern aspects of epidemiological surveillance of gonococcal infection spread

“…-об использовании метода NG-MAST для по-лучения генетической характеристики штаммов, ре-зистентных к антимикробным препаратам [103][104][105][106].…”

The nongenomic effects of progesterone in repressing iNOS activation through P38MAPK pathways in gonococci-infected polymorphonuclear leukocytes and the clinical significance

“…The strains were primary cultured. Genotyping of all positive isolates was performed by NG-MAST [1,10,14] . Strains were inoculated onto gonococcus (GC) agar plates or Luria-Bertani agar plates one night before the experiment started, suspended in GC broth (Difco) and grown to mid-logarithmic phase with 1% Kellogg's supplement (GCK), 1 µg/mL chloramphenicol or 2 µg/mL erythromycin.…”