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Catheter ablation of anteroseptal atrioventricular (AV) accessory pathway (AP) is still challenging because of their proximity to the normal AV conduction system that may be damaged with ablation. Traditionally, catheter ablation of anteroseptal APs via the inferior vena cava approach. In a few cases, anteroseptal AP ablation may fail because of conservative energy delivery at these sites or anatomical factor. In the recent years, a few cases of successful anteroseptal AP ablation in the noncoronary cusp (NCC) or sub tricuspid annulus have been reported. However, when need to map and ablate from the NCC or sub tricuspid annulus in patients with anteroseptal APs and what will be the mapping results at the NCC in patients with anteroseptal APs successful ablated in the, right anteroseptal region (RAS) have not been well evaluated. The purpose of this study was to discuss the characteristics of surface ECG, anatomic consideration, electrophysiology, and the strategy of ablation of anteroseptal APs in order to increase the success rate and safety of radiofrequency catheter ablation.There were 55 consecutive patients (age 53 ± 11 years, 36 male) out of 2200 patients presenting with anteroseptal APs who underwent RF ablation at our centre between July 2006 and March 2013. On the basis of successful ablation location, these patients were divided into right anteroseptal region group (RAS group, through inferior vena cava approach), noncoronary cusp group (NCC group, through retrograde aorta approach) and sub tricuspid annulus group (Sub-TA group, throngh superior vena cava approach). The clinical characteristics, surface electrocardiogram (ECG), intracardiac electrogram findings, and response to ablation in these patients between three groups were analysed. The strategies for mapping and ablation of anteroseptal APs were discussed.Successful ablation were achieved in all the 55 patients with anteroseptal APs. The sites with successful ablation were located in the RAS, adjacent to the His bundle region, in 48 patients including in 9 patients whose initial ablation were unsuccessful in the NCC. In the 4 cases with successful ablation in the NCC, the initial attempt ablation in the NCC without trying in the RAS was performed in 1 cases in whom the decision to target the NCC was based on previous experience of V-A fusion pattern adjacent to the His bundle region suggestive of an NCC AP site. In 1 of the 4 patients, irrigated energy (30–40 W and infusion rate of 17–30 ml/min) was used to eliminated the AP in the NCC after failed ablation with non-irrigated energy. There were 3 cases underwent successful ablation at Sub-TA region by a superior approach from the right internal jugular vein. There were no important complications during perioperative period. All the patients underwent a period of 8 months to 7 years of follow-up by telephone or outpatient department. Four patents ablated in RAS region recurred. No recurrence in patients underwent successful ablation in NCC or Sub-TA region.The results of catheter ablation of anteroseptal APs in a large case series indicate that anteroseptal APs in most patients can be successful ablated in RAS region, but in a few patients, anteroseptal APs need to be ablated from the NCC or Sub-TA region. The pre-excitation characteristics have no meaningful differences among the RAS, NCC and Sub-TA group. The incidence of anteroseptal AP which had to be ablated from NCC is relatively low, so ablation from NCC was not a preferential approach. The VA fusion pattern in RAS during retrograde AP conduction may be helpful to differentiate or predict successful ablation from RAS, NCC or Sub-TA in most patients with anteroseptal AP.