MLPA
(Abstract release date: 05/19/16)
EHA Library. Toral Ibarra D. 06/09/16; 134809; PB1909
Dr. Dennisse Sharon Toral Ibarra
Contributions
Contributions
Abstract
Abstract: PB1909
Type: Publication Only
Background
Since their inclusion in IPSS-R in 2012, cytogenetic alterations play a decisive role in the prognosis of patients with myelodysplastic syndrome (MDS). These alterations may be identified through several methods; cytogenetic study, including karyotype and FISH, is the most widely used and proved. However, new molecular techniques for the detection of these alterations have appeared which may be equally effective and far less expensive.
Aims
This study evaluates the MLPA cost-effectiveness in detecting cytogenetic alterations comparing it directly to FISH as gold standard test and karyotype.
Methods
We prospectively collected 131 bone morrow mononuclear cells samples from 118 MDS patients from September 2011 to June 2015. DNA was isolated by salting out and Multiplex Ligation-dependent Probe Amplification (MLPA), MRC Holland, following manufacturer recommendations. We did cytogenetic studies (karyotype and FISH) by conventional techniques.Human material and clinical information was obtained with informed consent and approval of institutional ethics committee.
Results
We obtained MLPA data from 131 bone marrow samples from 118 patients with morphological diagnosis of MDS, examined in our Institution. Karyotype was performed in 104 samples, the technique failed in six and 21 were not sent. FISH was only performed in cases with with abnormal or dubious karyotype; MLPA was performed in 125 samples, in 6 cases we did not receive sample. Alterations were detected in 39.4%, 71.4% and 52% by kariotype, FISH and MLPA, respectively. Specificity of MLPA could be assessed by comparing it with karyotype. Instead we could analyze the sensitivity of MLPA comparing with the most sensitive standard technique, FISH. Forty results were included for sensibility analysis, 31 studies corresponded to new diagnosis and 9 to follow-up samples.Comparing MLPA with karyotype, MLPA showed a high specificity 95% (p<0.05) (S: 71% E 95%).We could compare fISH and MLPA in 40 cases (Figure), MLPA analysis showed sensitivity of 75% and specificity of 85.7% (p<0.05). The results were correlated in 74.2%. We observed a increase of sensitivity (85% and 82% (p<0.05) in the subgroup of new diagnosis. Moreover, MLPA detected several alterations which had gone unobserved by FISH, such as: –Y, SMAD4+ y 8pq+. MLPA also proved less expensive in both reagents and staff time.
Conclusion
Our data indicate that MLPA is a useful tool, as valid as FISH and with a better cost-effectiveness. Although it has sensitivity limitations (in cases of follow-up), it can be used as first-line screening and also as a complementary test for the cytogenetic study. Furthermore, MLPA allows the simultaneous assessment of many loci and the detail study of the most important ones, adding more information to the analysis of copy number variations, which are the most frequent and relevant abnormalities in MDS.
Session topic: E-poster
Type: Publication Only
Background
Since their inclusion in IPSS-R in 2012, cytogenetic alterations play a decisive role in the prognosis of patients with myelodysplastic syndrome (MDS). These alterations may be identified through several methods; cytogenetic study, including karyotype and FISH, is the most widely used and proved. However, new molecular techniques for the detection of these alterations have appeared which may be equally effective and far less expensive.
Aims
This study evaluates the MLPA cost-effectiveness in detecting cytogenetic alterations comparing it directly to FISH as gold standard test and karyotype.
Methods
We prospectively collected 131 bone morrow mononuclear cells samples from 118 MDS patients from September 2011 to June 2015. DNA was isolated by salting out and Multiplex Ligation-dependent Probe Amplification (MLPA), MRC Holland, following manufacturer recommendations. We did cytogenetic studies (karyotype and FISH) by conventional techniques.Human material and clinical information was obtained with informed consent and approval of institutional ethics committee.
Results
We obtained MLPA data from 131 bone marrow samples from 118 patients with morphological diagnosis of MDS, examined in our Institution. Karyotype was performed in 104 samples, the technique failed in six and 21 were not sent. FISH was only performed in cases with with abnormal or dubious karyotype; MLPA was performed in 125 samples, in 6 cases we did not receive sample. Alterations were detected in 39.4%, 71.4% and 52% by kariotype, FISH and MLPA, respectively. Specificity of MLPA could be assessed by comparing it with karyotype. Instead we could analyze the sensitivity of MLPA comparing with the most sensitive standard technique, FISH. Forty results were included for sensibility analysis, 31 studies corresponded to new diagnosis and 9 to follow-up samples.Comparing MLPA with karyotype, MLPA showed a high specificity 95% (p<0.05) (S: 71% E 95%).We could compare fISH and MLPA in 40 cases (Figure), MLPA analysis showed sensitivity of 75% and specificity of 85.7% (p<0.05). The results were correlated in 74.2%. We observed a increase of sensitivity (85% and 82% (p<0.05) in the subgroup of new diagnosis. Moreover, MLPA detected several alterations which had gone unobserved by FISH, such as: –Y, SMAD4+ y 8pq+. MLPA also proved less expensive in both reagents and staff time.
Conclusion
Our data indicate that MLPA is a useful tool, as valid as FISH and with a better cost-effectiveness. Although it has sensitivity limitations (in cases of follow-up), it can be used as first-line screening and also as a complementary test for the cytogenetic study. Furthermore, MLPA allows the simultaneous assessment of many loci and the detail study of the most important ones, adding more information to the analysis of copy number variations, which are the most frequent and relevant abnormalities in MDS.
Session topic: E-poster
Abstract: PB1909
Type: Publication Only
Background
Since their inclusion in IPSS-R in 2012, cytogenetic alterations play a decisive role in the prognosis of patients with myelodysplastic syndrome (MDS). These alterations may be identified through several methods; cytogenetic study, including karyotype and FISH, is the most widely used and proved. However, new molecular techniques for the detection of these alterations have appeared which may be equally effective and far less expensive.
Aims
This study evaluates the MLPA cost-effectiveness in detecting cytogenetic alterations comparing it directly to FISH as gold standard test and karyotype.
Methods
We prospectively collected 131 bone morrow mononuclear cells samples from 118 MDS patients from September 2011 to June 2015. DNA was isolated by salting out and Multiplex Ligation-dependent Probe Amplification (MLPA), MRC Holland, following manufacturer recommendations. We did cytogenetic studies (karyotype and FISH) by conventional techniques.Human material and clinical information was obtained with informed consent and approval of institutional ethics committee.
Results
We obtained MLPA data from 131 bone marrow samples from 118 patients with morphological diagnosis of MDS, examined in our Institution. Karyotype was performed in 104 samples, the technique failed in six and 21 were not sent. FISH was only performed in cases with with abnormal or dubious karyotype; MLPA was performed in 125 samples, in 6 cases we did not receive sample. Alterations were detected in 39.4%, 71.4% and 52% by kariotype, FISH and MLPA, respectively. Specificity of MLPA could be assessed by comparing it with karyotype. Instead we could analyze the sensitivity of MLPA comparing with the most sensitive standard technique, FISH. Forty results were included for sensibility analysis, 31 studies corresponded to new diagnosis and 9 to follow-up samples.Comparing MLPA with karyotype, MLPA showed a high specificity 95% (p<0.05) (S: 71% E 95%).We could compare fISH and MLPA in 40 cases (Figure), MLPA analysis showed sensitivity of 75% and specificity of 85.7% (p<0.05). The results were correlated in 74.2%. We observed a increase of sensitivity (85% and 82% (p<0.05) in the subgroup of new diagnosis. Moreover, MLPA detected several alterations which had gone unobserved by FISH, such as: –Y, SMAD4+ y 8pq+. MLPA also proved less expensive in both reagents and staff time.
Conclusion
Our data indicate that MLPA is a useful tool, as valid as FISH and with a better cost-effectiveness. Although it has sensitivity limitations (in cases of follow-up), it can be used as first-line screening and also as a complementary test for the cytogenetic study. Furthermore, MLPA allows the simultaneous assessment of many loci and the detail study of the most important ones, adding more information to the analysis of copy number variations, which are the most frequent and relevant abnormalities in MDS.
Session topic: E-poster
Type: Publication Only
Background
Since their inclusion in IPSS-R in 2012, cytogenetic alterations play a decisive role in the prognosis of patients with myelodysplastic syndrome (MDS). These alterations may be identified through several methods; cytogenetic study, including karyotype and FISH, is the most widely used and proved. However, new molecular techniques for the detection of these alterations have appeared which may be equally effective and far less expensive.
Aims
This study evaluates the MLPA cost-effectiveness in detecting cytogenetic alterations comparing it directly to FISH as gold standard test and karyotype.
Methods
We prospectively collected 131 bone morrow mononuclear cells samples from 118 MDS patients from September 2011 to June 2015. DNA was isolated by salting out and Multiplex Ligation-dependent Probe Amplification (MLPA), MRC Holland, following manufacturer recommendations. We did cytogenetic studies (karyotype and FISH) by conventional techniques.Human material and clinical information was obtained with informed consent and approval of institutional ethics committee.
Results
We obtained MLPA data from 131 bone marrow samples from 118 patients with morphological diagnosis of MDS, examined in our Institution. Karyotype was performed in 104 samples, the technique failed in six and 21 were not sent. FISH was only performed in cases with with abnormal or dubious karyotype; MLPA was performed in 125 samples, in 6 cases we did not receive sample. Alterations were detected in 39.4%, 71.4% and 52% by kariotype, FISH and MLPA, respectively. Specificity of MLPA could be assessed by comparing it with karyotype. Instead we could analyze the sensitivity of MLPA comparing with the most sensitive standard technique, FISH. Forty results were included for sensibility analysis, 31 studies corresponded to new diagnosis and 9 to follow-up samples.Comparing MLPA with karyotype, MLPA showed a high specificity 95% (p<0.05) (S: 71% E 95%).We could compare fISH and MLPA in 40 cases (Figure), MLPA analysis showed sensitivity of 75% and specificity of 85.7% (p<0.05). The results were correlated in 74.2%. We observed a increase of sensitivity (85% and 82% (p<0.05) in the subgroup of new diagnosis. Moreover, MLPA detected several alterations which had gone unobserved by FISH, such as: –Y, SMAD4+ y 8pq+. MLPA also proved less expensive in both reagents and staff time.
Conclusion
Our data indicate that MLPA is a useful tool, as valid as FISH and with a better cost-effectiveness. Although it has sensitivity limitations (in cases of follow-up), it can be used as first-line screening and also as a complementary test for the cytogenetic study. Furthermore, MLPA allows the simultaneous assessment of many loci and the detail study of the most important ones, adding more information to the analysis of copy number variations, which are the most frequent and relevant abnormalities in MDS.
Session topic: E-poster
{{ help_message }}
{{filter}}