Accurate and reproducible quantification of right ventricular volumes and function plays a crucial role in the diagnosis of various cardiac diseases. It also guides clinical decision-making and monitoring of therapy in many conditions. These include pathologies primarily affecting the right ventricle (RV) such as arrhythmogenic right ventricular cardiomyopathy (ARVC), tricuspid regurgitation, pulmonary hypertension as well as congenital heart disease (CHD) entities such as Tetralogy of Fallot (ToF), Ebstein’s and shunt diseases [1–6].

Various studies have established cardiovascular magnetic resonance (CMR) as the standard of reference in assessment of RV dimensions and function [7–10]. Furthermore, accurate quantification of RV dimensions has gained attention as an important predictor of outcomes in heart failure and proven essential for therapy decisions and surveillance [5–8, 11, 12] Other than left ventricular assessment the complex anatomy of the RV imposes a challenge to other volumetric measurement methods with additional limitations of echocardiography related to limited acoustic windows [4, 13]. This may be further complicated by various RV morphologies in CHD.

While the acquisition of cine steady state free prcession (SSFP) data in short axis orientation (SAO) is the accepted standard for assessment of the left ventricle (LV) in CMR, both, transverse as well as short axis slice orientations have been applied for analysis of the RV. However, acquisition of an additional transverse stack for the sole purpose of RV volumetric assessment not only lengthens the total examination time and as such effects imaging workflow and capcities but also may negatively impact patients’ comfort and compliance.

Several studies have demonstrated that transverse cine orientation has better intra- and inter-observer reproducibility than SAO cine in patient populations with various anatomies [7, 14–17].

Additionally, longitudinal follow-up examinations using CMR and identification of changes and pattern of changes over time have gained attention especially in CHD [12, 18]. CMR has previously demonstrated high test-retest reproducibilities for the assessment of RV and LV volumes with superior results in comparison to echocardiography [19–21]. While few studies have aimed for the test-retest reproducibility with respect to LV and RV size and function using SAO slices [19, 22–25], no study has yet directly compared the influence of slice orientation on test-retest reproducibility for RV assessment. Therefore it still remains unclear which slice orientation provides better overall performance including test-retest reproducibility for longitudinal follow-up of RV volumes and ejection fraction (EF).

We hypothesize that SAO is not inferior to transverse orientation cine SSFP in respect to the overall variability of longitudinal comparison for RV volume and function assessment.

The aim of our study therefore was to evaluate test-retest variability of cine SSFP applied in both orientations approaches in addition to intra- and inter-observer variability in a cohort of healthy volunteers undergoing two serial CMR exams.

Evaluation of RV volumes and function based on cine SSFP was successful in the entire study cohort of 18 healthy volunteers with a total of 36 cine SSFP volume stacks to be contoured (18 transverse, 18 short axis). In all but one volunteer MPA PC flow assessment was successfully analysed for both sessions. A single volunteer demonstrated a significant variation in RR interval (1039 ± 422 ms) in one session of the MPA PC flow data related to transient arrhythmia making this flow measurement unreliable as a reference for the RV output. This volunteer’s MPA flow data was therefore not used for any comparison of flow related data between both measurement sessions. All other flow and volumetric data of this individual were not affected by any arrhythmia and as such included in respective subsequent data analysis.

In the 17 volunteers where reliable heart rate recording was available, no significant differences between the recorded heart rates between scan session 1 and 2 was observed (65.5 ± 7.4 bpm vs. 67.3 ± 10.2 bpm: P = 0.12).

Main pulmonary artery flow and RV stroke volume

Quantification of the MPA flow for measurement sessions 1 and 2 demonstrated strong correlation between both sessions (r = 0.959; P < 0.001) without any significant difference in forward flow (Table 1) (Fig. 2). The resulting RV output per minute averaged to 5948.2 ± 1189.1 ml/min (3876.0–7936.1 ml) for session 1 and 5717.5 ± 1098.7 ml/min (3842.4–7617.6 ml) for session 2 (P = 0.1).

	Observer 1		Observer 2		Observer 3
	Transverse	Short axis	Transverse	Short axis	MPA PC flow
Scan 1 (n = 17)	92.5 ± 15.8 (66.9–123.3)	86.9 ± 14.5 (64.0–114.6)	92.0 ± 15.4 (60.1–121.2)	84.9 ± 15.6 (56.9–114.5)	89.8 ± 16.6 (67.7–118.7)
Scan 2 (n = 18)	90.3 ± 14.1 (68.3–116.8)	87.1 ± 15.1 (61.3–116.3)	89.2 ± 14.2 (60.6–114.0)	84.9 ± 13.7 (61.8–111.3)	87.2 ± 14.9 (66.6–114.4)

All data in [ml]; data is presented in mean ± standard deviation; range in parentheses

No significant differences were observed for RV SV results of both observers and both imaging orientations in comparison to the respective MPA forward volumes as measured by PC flow measurements (all P > 0.05) (Table 1). In addition RV SV results of both observers demonstrated strong positive correlation to independently evaluated MPA flow data for transverse orientation (observer 1: r = 0.896; observer 2: r = 0.873; all P < 0.001) and SAO (observer 1: r = 0.826; observer 2: r = 0.824; all P < 0.001). For both observers the transverse orientation RV SV strongly correlated to SAO derived RV SV data (observer 1: r = 0.845; observer 2: r = 0.924; all P < 0.001) without significant differences in pairwise comparison (all P > 0.05) (Table 2)

	Scan session 1				Scan session 2
	Observer 1		Observer 2		Observer 1		Observer 2
	Transverse	Short axis	Transverse	Short axis	Transverse	Short axis	Transverse	Short axis
EDV (ml)	163.2 ± 26.5 (117.2–212.6)	160.5 ± 26.2 (117.7–215.1)	164.9 ± 25.5 (116.3–215.9)	157.8 ± 27.6 (106.5–215.5)	161.4 ± 25.9 (112.3–220.0)	160.2 ± 28.5 (109.1–223.1)	163.0 ± 26.8 (113.2–224.0)	157.3 ± 25.8 (111.6–214.4)
ESV (ml)	71.3 ± 13.6 (49.3–96.5)	74.4 ± 14.7 (46.3–105.9)	73.5 ± 13.0 (56.2–104.5)	73.6 ± 14.3 (47.5–105.5)	71.2 ± 15.4 (43.9–103.1)	73.1 ± 15.6 (45.6–107.1)	73.9 ± 14.9 (50.1–110.0)	72.3 ± 14.3 (49.8–103.2
SV (ml)	91.9 ± 15.6 (66.9–123.3)	86.0 ± 14.6 (64.0–114.6)	91.4 ± 15.1(60.1–121.2)	84.1 ± 15.4 (56.9–114.5)	90.3 ± 14.1 (68.3–116.8)	87.1 ± 15.1 (61.3–116.3)	89.2 ± 14.2 (60.6–114.0)	84.9 ± 13.7 (61.8–111.3)
EF (%)	56.4 ± 3.5 (49.9–60.4)	53.8 ± 4.1 (47.3–60.9)	55.4 ± 3.5 (50.2–62.9)	53.4 ± 3.4 (46.6–60.3)	56.1 ± 4.9 (47.8–64.5)	54.5 ± 3.8 (47.8–60.9)	54.8 ± 3.7 (47.5–61.4)	54.1 ± 3.7 (46.0–60.2)

Data is presented in mean ± standard deviation; range in parentheses * P < 0.05

For inter-observer agreement the ICC demonstrated higher values in short-axis vs. transverse orientation for SV measures (0.954 [0.882, 0.983] vs. 0.936 [0.837, 0.976]) (Table 3).

	Transverse	Short axis
RV EDV	0.983 (0.954, 0.993)	0.986 (0.963, 0.995)
RV ESV	0.926 (0.814, 0.972)	0.955 (0.883, 0.983)
RV SV	0.936 (0.837, 0.976)	0.954 (0.882, 0.983)
RV EF	0.625 (0.238, 0.841)	0.769 (0.482, 0.907)

ICC values with 95 % CI in parentheses

No significant interaction between imaging orientation was demonstrated for interstudy (F = 0.402, p = 0.527), inter-observer (F = 0.143, p = 0.706) and intra-observer (F = 0.095, p = 0.759) variability of SV results.

End-diastolic volumes

Contouring of both observers did not demonstrate any significant differences between short axis and transverse cine SSFP for EDV (Table 2). EDV results based on SAO demonstrated strong correlation to those based on transverse slice orientation (observer 1: r = 0.952; observer 2: r = 0.982; all P < 0.001). Results for EDV demonstrated a bias of 0.4 % (−9.5 %,10.3 %) for short axis and 1.1 % (−7.3 %,9.4 %) for transverse orientation between both sessions (Table 4) (Fig. 3). In regard to inter- and intra-observer variability SAO demonstrated a bias of 1.9 % (−4.8 %, 8.7 %) and 0.5 % (−5.1 %, 6.0 %) while for transverse slice orientation the bias was −1.1 % (−7.1 %, 4.8 %) and 0.3 %, 7.7 %), respectively (Table 4) (Fig. 3). For inter-observer agreement the ICC demonstrated minimally higher values in short-axis vs. transverse orientation for EDV (0.986 [0.963,0.995] vs. 0.983 [0.954,0.993]) but overlapping confidence intervals (Table 3).

	Transverse	Short axis
EDV
Interstudy	1.1 (−7.3, 9.4)	0.4 (−9.5, 10.3)
Inter-observer	−1.1 (−7.1, 4.8)	1.9 (−4.8, 8.7)
Intra-observer	0.3 (−7.0, 7.7)	0.5 (−5.1, 6.0)
ESV
Interstudy	0.8 (−16.0, 17.6)	2.1 (−12.3, 16.4)
Inter-observer	−3.2 (−17.6, 11.2)	1.0 (−11.2, 13.2)
Intra-observer	−1.4 (−14.5, 11.7)	−0.4 (−12.4, 11.6)
EF
Interstudy	0.6 (−11.2, 12.4)	−1.5 (−7.2, 10.2)
Inter-observer	1.6 (−9.1, 12.4)	0.5 (−8.7, 9.7)
Intra-observer	1.6 (−7.2, 10.5)	0.7 (−8.4, 9.7)

Values demonstrate % bias with BA limits of agreement in parentheses

2-way ANOVA did not demonstrate significant interaction between imaging orientation and interstudy (F = 0.050; p = 0.824) as well as inter- (F = 0.382; p = 0.537) and intra-observer (F = 0.123; p = 0.726) variability for EDV results.

End-systolic volumes

No significant differences were identified with respect to ESV data for both observers between short axis and transverse cine SSFP orientation (Table 2). ESV results based on SAO demonstrated strong correlation to those based on transverse slice orientation (observer 1: r = 0.925; observer 2: r = 0.904; all P < 0.001). Bland-Altman analysis revealed a bias of 2.1 % (−12.3 %, 16.4 %) and 0.8 % (−16.0 %, 17.6 %) between both studies for short axis and transverse orientation, respectively (Table 4) (Fig. 4). In regard to inter- and intra-observer variability of ESV data SAO demonstrated a bias of 1.0 % (−11.2 %, 13.2 %) and −0.4 % (−12.4 %, 11.6 %) while for transverse slice orientation the bias was −3.2 % (−17.6 %, 11.2 %) and −1.4 % (−14.5 %, 11.7 %), respectively (Table 4) (Fig. 4). For inter-observer agreement the ICC also demonstrated higher values in short-axis vs. transverse orientation for ESV (0.955 [0.883, 0.983] vs. 0.926 [0.814, 0.972]) with again overlapping confidence intervals (Table 3).

No significant interaction between imaging orientation was demonstrated for interstudy (F = 0.069, p = 0.792), inter-observer (F = 0.581, p = 0.447) and intra-observer (F = 0.119, p = 0.730) variability of ESV results.

Ejection fraction

Resulting RV EF did not demonstate any significant differences between short axis and transverse cine SSFP orientation for the two observers (Table 2). The inter-observer agreement for RV ejection fraction as assessed by ICC was lower than for individually measured volumes (e.g., EDV, ESV) (Table 3). It was again higher for SAO (0.769 [0.482, 0.907]) than for transverse orientation (0.625 [0.238, 0.841] (Table 3). The bias for EF measurements between both studies as assessed by Bland-Altman plots was −1.5 % (−7.2 %, 10.2 %) for a stack of short axis slices in comparison to 0.6 % (−11.2 %, 12.4 %) when using a stack of transverse cine SSFP slices for RV volumetric evaluation (Table 4) (Fig. 5). For inter-observer variability the bias was 0.5 % (−8.7 %, 9.7 %) for short axis and 1.6 % (−9.1 %, 12.4 %) for transverse orientation and for intra-observer variability the bias was 0.7 % (−8.4 %, 9.7 %) and 1.6 % (−7.2 %, 10.5 %), respectively (Table 4) (Fig. 5).

Again, no significant interaction between imaging orientation was demonstrated for interstudy (F = 0.726, p = 0.395), inter-observer (F = 0.338, p = 0.561) and intra-observer (F = 0.031, p = 0.862) variability of ESV results.

This study demonstrates that there is no significant effect of the slice orientation on accuracy or intra-observer and inter-observer variability of volumetric and functional assessment of the right ventricle in a cohort with normal RV morphology and size. Furthermore, it establishes that in this cohort there is no significant difference in test-retest-reproducibility between both slice orientations. While previous studies have focused on the effects of slice orientation on intra- and interobserver variability in RV volumetric analysis, to the best of our knowledge this study presents the first report on test-retest reproducibility comparison of short axis and transverse slice orientations cine CMR based RV volumetric assessment.

While for the acquisition of RV volumetric data either transverse or short axis slice orientations have been recommended for cine SSFP [27], published normal values for RV size and function are almost exclusively based on SAO and allow further subcategorization for gender and age [14, 28–31]. As such there is limited available published normal data of RV volumetry on transverse slice orientation.

Various investigators have demonstrated significantly lower intra- and inter-observer variability for RV volume assessment based on transverse over short axis orientation for cine SSFP examinations. Alfakih et al. demonstrated better intra- and inter-observer reproducibility for transverse orientation in healthy volunteers, while Fratz et al. demonstrated significantly lower intra- and inter-observer variance for RV (and LV) volumetry utilizing transverse cine SSFP orientation in a population of repaired tetralogy of Fallot (rToF) patients [16]. Similar results have been demonstrated in patients with atrial redirection surgery for complete transposition of the great arteries as well as in patients with Ebstein anomaly [17, 32]. Other studies showed a similar trend but no clinical significance of findings. Clarke et al. categorized differences in intra- and inter-observer variability between both orientations as not clinically relevant [14, 15]. Applying PC flow measurements as an independent standard in our population, the results of this study confirms that slice orientation has no significant impact on either intra- nor inter-observer variability in assessment of RV volumes and global systolic function (Tables 2 and 4).

In addition to intra- and inter-observer variability few studies have also demonstrated differences in ventricular volumes between both slice orientations. In the work by Alfakih et al. short axis slice orientations resulted in significantly larger results for RV EDV (4.7 % difference) and RV ESV (10.7 % difference) while no differences were found for SV [7]. With 2.9 % lower RV EDV values and 5.8 % lower RV ESV values derived from short axis slice orientation Sarikouch et al. reported lower off-sets in healthy children and adolescents (8-20y); again no relevant relative differences were demonstrated for resulting SV (1.1 % lower) and EF (1.8 % higher) using short axis orinentation [33]. Fratz et al. demonstrated significant differences in volumes for both approaches in rToF patients, while Clarke et al. showed a small, though statistically significant difference between both slice orientations for RV ESV but no significant differences for overall RV size as measured by EDV in various types of CHD (excluding subaortic RV) [14, 16]. Results published by James et al. in fact demonstrated minimaly lower RV EDV when applying short axis slice orientation with no significant EF differences; in addition they applied PC flow measurements in the MPA as an independent standard of reference and demonstrated no differences in RV SV offsets to PC data [15]. With previous studies demonstrating inconsistent results with either overestimation or underestimation of RV size by short axis cine SSFP, our study confirms that with regard to RV volumes no differences exist in both orientations. While underlying reasons for differences in previous studies are hard to evaluate, inconsistency in contouring approaches might have played a role. In addition, transverse orientation cine add extra examination time and are more vulnerable to errors introduced by inconsistent breath-holding than SAO cines (related to the parallel orientation of the inferior RV boundaries to the slice orientation). In unwell patients longer scans result in greater fatigue and may increase inconsistency; as such this is more likely to result in physiological changes possibly resulting in misinterpretation as significant volumetric findings (e.g., differences in heart rate during acquisitions for both orientations may be misinterpreted as discrepancy between RV and LV stroke volumes) or may introduce errors into other calculations (e.g., volume assessment of valvular heart disease or intracardiac shunts).

In therapy guidance and longitudinal follow-up of various cardiac pathologies that affect ventricular volumes and function a high test-retest reproducibility is of utmost importance. For example, various investigators have proposed cut-off values for timing of pulmonary valve replacement (PVR) in patients with repaired tetralogy of Fallot and chronic pulmonary valve regurgitation. With an RV EDV of <160–165 ml/m² or an RV ESV of <80–85 ml/m² a normalization of RV size after PVR has been demonstrated [3, 5, 6]. Grotheus et al. have demonstrated the substantial impact of the high test-retest reproducibility of CMR (compared to echocardiography) in assessment of LV volumes and function with a much more sensitive identification of true changes in ventricular size and performance [25]. Few studies have also assessed interstudy reproducibility for RV parameters. Early investigations by Pattynama et al. with 40 consecutive scans in 2 volunteers over 6 weeks have demonstrated a large variability of results from scan to scan using SAO [23]. However, the applied techniques (spin-echo and free-breathing spoiled gradient echo) are no longer in use for cardiac functional imaging and as such the presented data does likely not reflect current imaging approaches [31]. Using breath-hold FLASH techniques in SAO Grothues et al. demonstrated low interstudy variability of RV parameters healthy subjects, patients with heart failure, and patients with LV hypertrophy [19]. They reported an RV interstudy reproducibility (range between groups) of 6.2 % (4.2 %–7.8 %) for EDV and 14.1 % (8.1 %–18.1 %) for ESV [19]. Most recently Blalock et al. again investigated a population with rTOF demonstrating a repeatability coefficient (2 SD of the differences; percent value of population mean) of 9 % for RV EDV and 13 % for RV ESV when the same observer contoured both examinations [24]. While no change in results was demonstrated for RV EDV when a second obsverver contoured the repeat study, the repeatability coefficient for RV ESV increased to 20 % [24]. While all above referenced studies on test-retest performance for RV size and function focused on SAO, no data is available for either transverse slice selection nor for a comparison of both approaches. Interpretation of our data confirms a high test-retest reproducibility of CMR in assessment of RV volumes and size which is independent of the slice orientation chosen for ventricular coverage. Calculation of the minimal detectable difference between consecutive examinations based on standard error of the measurement (SEM) computation demonstrate that for RV EDV both orientations can identify real changes as small as ~10-12 ml, while for RV EDV identifiable changes are even smaller with ~8–9 ml between studies. This data results in identification of real interstudy RV EF changes of as small as ~4–5 %.

Despite the fact that this study provides the first direct comparison of test-retest reproducibility between transverse and short axis slice orientation for RV volumes and function, the following limitations apply.

The study is composed on a relatively small number of individuals included, however as opposed to previous test-retest investigations we included MPA PC flow measurements as an independent reference standard. Furthermore, our study only recruited cooperative healthy volunteers without relevant arrhythmia and a high level of compliance to scanning instructions. In a symptomatic patient population the test-retest reproducibility therefore might be lower than reported above.

This first report on direct comparison of short axis and transverse cine SSFP slice orientation with respect to test-retest reproducibility of RV volumes and function demonstrate equally high performance of both orientations. While there were comparable levels of intra- and interobserver variability and similar volumetric results of both slice orientations, volumetric analysis of transverse slice orientation cines did not demonstrate a significant advantage over short axis ventricular coverage. With standard coverage using SAO for LV assessment, this single data set also provides adequate assessment of RV size and function. As such, transverse cine SSFP imaging may be omitted for the sole purpose of RV volumetric assessment resulting in shortened scan protocols for improved workflow and patient comfort.

However, assessment of complex vascular and cardiac anatomy may still require the application of transverse cine SSFP as a non-oblique and reproducible orientation for insight into spatial relationships.