Ben Salem Y, Boullegue O, Mastouri M, Ktata S, Boujaafar N, Mzoughi R
Faculté de Pharmacie de Monastir, Tunisie.
Pathol Biol (Paris). 2006 Apr;54(3):137-47. doi: 10.1016/j.patbio.2005.04.004. Epub 2005 Jun 17.
We reported a molecular characterization of 25 Haemophilus influenzae strains derived from cases of meningitis and sepsis in children aged less than five years hospitalized in pediatric wards from three hospitals in the Sahel area (Tunisia) during the period 1997-2002. These strains were biotyped and subjected to a capsular typing by Slide agglutination serotyping and Polymerase Chain Reaction (PCR). The genetic polymorphism of these strains was also studied in Arbitrarily Primed Polymerase Chain Reaction (AP-PCR) with two sets of primers: RAP IV and 217 delta(2) as in Pulsed Field Gel Electrophoresis after digestion of the total DNA with the restriction enzyme SmaI (PFGE SmaI). Nineteen strains among 25 (76%) were of biotype I. The bexA gene was highlighted in 13 strains (52%) and in all the cases it was of the type b. Twelve strains (48%) were shown to be unencapsulated by PCR. AP-PCR RAP IV (23 genotypes/25 with a discrimination index ID=0.993) had shown nearly the same discriminatory power than PFGE (20 genotypes/21 strains with a discrimination index ID=0.995). We thus note, how capsular typing by PCR is more sensitive than slide agglutination serotyping. We also note the genetic diversity of the invasive strains isolated with a remarkable presence of non typable strains. AP PCR seems to be an alternative of choice for the epidemiologic follow-up of the Haemophilus influenzae invasive infections.
我们报告了1997年至2002年期间,从萨赫勒地区(突尼斯)三家医院儿科病房收治的5岁以下儿童脑膜炎和败血症病例中分离出的25株流感嗜血杆菌的分子特征。这些菌株进行了生物分型,并通过玻片凝集血清分型和聚合酶链反应(PCR)进行荚膜分型。还使用两组引物:RAP IV和217 delta(2),通过任意引物聚合酶链反应(AP-PCR)研究了这些菌株的遗传多态性,如同用限制性内切酶SmaI消化总DNA后的脉冲场凝胶电泳(PFGE SmaI)。25株菌株中有19株(76%)为生物I型。13株菌株(52%)中检测到bexA基因,且所有病例中该基因均为b型。PCR结果显示12株菌株(48%)无荚膜。AP-PCR RAP IV(25株中有23种基因型,鉴别指数ID = 0.993)显示出与PFGE(21株中有20种基因型,鉴别指数ID = 0.995)几乎相同的鉴别能力。因此我们注意到,PCR荚膜分型比玻片凝集血清分型更敏感。我们还注意到分离出的侵袭性菌株的遗传多样性,其中不可分型菌株显著存在。AP-PCR似乎是流感嗜血杆菌侵袭性感染流行病学随访的一种首选替代方法。