IMPROVED DETECTION OF CHLAMYDOPHILA PSITTACI DNA IN OCULAR ADNEXAL MALT LYMPHOMA WITH PCR OPTIMIZATION
(Abstract release date: 05/21/15)
EHA Library. Yoo C. 06/12/15; 102708; PB1794
Disclosure(s): Asan Medical Center, University of Ulsan College of MedicineOncology
Changhoon Yoo
Contributions
Contributions
Abstract
Abstract: PB1794
Type: Publication Only
Background
The pathogenic association between Chlamydophila psittaci (Cp) infection and ocular adnexal MALT lymphoma (OAML) has been suggested, but large geographic variation (0-87%) has been reported in the prevalence of Cp infection in patients with OAML. Although its cause has not been elucidated fully, this geographic discrepancy may be in part due to the methodological biases or suboptimal PCR.
Aims
Therefore, we examined the detection rates for Cp DNA in patients with OAML according to the different PCR condition.
Methods
Six OAML patients diagnosed between April 8, 2014 and August 11, 2014 were included in this study. DNA was extracted from paraffin-embedded tissues and touchdown enzyme time release (TETR)-PCR was performed to identify Cp DNA as previously reported (Madico et al., J Clin Microbiol 2000;38:1085-93). Then, we performed TETR-PCR with modified annealing temperature (beginning at 67C vs 62C in original protocol) using same extracted DNA samples.
Results
Four (67%) patients were male and median age was 50 years (range, 41-77 years). The primary site of OAML was conjunctiva in 4 (67%) patients and orbital soft tissue in 2 (33%) patients. At presentation, each 3 (50%) patients were diagnosed as stage I and IV, respectively. In initial TETR-PCR according to the original protocol, Cp DNA was detected in none of 6 patients. When TETR-PCR was performed with modification of annealing temperature, Cp DNA was identified in 4 (67%) patients. Direct sequencing could be performed in 3 of 4 positive cases, and specificity of amplified fragments was confirmed.
Summary
PCR optimization with modification of annealing temperature could improve the detection of Cp DNA in patients with OAML. Future prospective validation in a larger sample size is necessary.
Keyword(s): Infection, MALT lymphoma
Type: Publication Only
Background
The pathogenic association between Chlamydophila psittaci (Cp) infection and ocular adnexal MALT lymphoma (OAML) has been suggested, but large geographic variation (0-87%) has been reported in the prevalence of Cp infection in patients with OAML. Although its cause has not been elucidated fully, this geographic discrepancy may be in part due to the methodological biases or suboptimal PCR.
Aims
Therefore, we examined the detection rates for Cp DNA in patients with OAML according to the different PCR condition.
Methods
Six OAML patients diagnosed between April 8, 2014 and August 11, 2014 were included in this study. DNA was extracted from paraffin-embedded tissues and touchdown enzyme time release (TETR)-PCR was performed to identify Cp DNA as previously reported (Madico et al., J Clin Microbiol 2000;38:1085-93). Then, we performed TETR-PCR with modified annealing temperature (beginning at 67C vs 62C in original protocol) using same extracted DNA samples.
Results
Four (67%) patients were male and median age was 50 years (range, 41-77 years). The primary site of OAML was conjunctiva in 4 (67%) patients and orbital soft tissue in 2 (33%) patients. At presentation, each 3 (50%) patients were diagnosed as stage I and IV, respectively. In initial TETR-PCR according to the original protocol, Cp DNA was detected in none of 6 patients. When TETR-PCR was performed with modification of annealing temperature, Cp DNA was identified in 4 (67%) patients. Direct sequencing could be performed in 3 of 4 positive cases, and specificity of amplified fragments was confirmed.
Summary
PCR optimization with modification of annealing temperature could improve the detection of Cp DNA in patients with OAML. Future prospective validation in a larger sample size is necessary.
Keyword(s): Infection, MALT lymphoma
Abstract: PB1794
Type: Publication Only
Background
The pathogenic association between Chlamydophila psittaci (Cp) infection and ocular adnexal MALT lymphoma (OAML) has been suggested, but large geographic variation (0-87%) has been reported in the prevalence of Cp infection in patients with OAML. Although its cause has not been elucidated fully, this geographic discrepancy may be in part due to the methodological biases or suboptimal PCR.
Aims
Therefore, we examined the detection rates for Cp DNA in patients with OAML according to the different PCR condition.
Methods
Six OAML patients diagnosed between April 8, 2014 and August 11, 2014 were included in this study. DNA was extracted from paraffin-embedded tissues and touchdown enzyme time release (TETR)-PCR was performed to identify Cp DNA as previously reported (Madico et al., J Clin Microbiol 2000;38:1085-93). Then, we performed TETR-PCR with modified annealing temperature (beginning at 67C vs 62C in original protocol) using same extracted DNA samples.
Results
Four (67%) patients were male and median age was 50 years (range, 41-77 years). The primary site of OAML was conjunctiva in 4 (67%) patients and orbital soft tissue in 2 (33%) patients. At presentation, each 3 (50%) patients were diagnosed as stage I and IV, respectively. In initial TETR-PCR according to the original protocol, Cp DNA was detected in none of 6 patients. When TETR-PCR was performed with modification of annealing temperature, Cp DNA was identified in 4 (67%) patients. Direct sequencing could be performed in 3 of 4 positive cases, and specificity of amplified fragments was confirmed.
Summary
PCR optimization with modification of annealing temperature could improve the detection of Cp DNA in patients with OAML. Future prospective validation in a larger sample size is necessary.
Keyword(s): Infection, MALT lymphoma
Type: Publication Only
Background
The pathogenic association between Chlamydophila psittaci (Cp) infection and ocular adnexal MALT lymphoma (OAML) has been suggested, but large geographic variation (0-87%) has been reported in the prevalence of Cp infection in patients with OAML. Although its cause has not been elucidated fully, this geographic discrepancy may be in part due to the methodological biases or suboptimal PCR.
Aims
Therefore, we examined the detection rates for Cp DNA in patients with OAML according to the different PCR condition.
Methods
Six OAML patients diagnosed between April 8, 2014 and August 11, 2014 were included in this study. DNA was extracted from paraffin-embedded tissues and touchdown enzyme time release (TETR)-PCR was performed to identify Cp DNA as previously reported (Madico et al., J Clin Microbiol 2000;38:1085-93). Then, we performed TETR-PCR with modified annealing temperature (beginning at 67C vs 62C in original protocol) using same extracted DNA samples.
Results
Four (67%) patients were male and median age was 50 years (range, 41-77 years). The primary site of OAML was conjunctiva in 4 (67%) patients and orbital soft tissue in 2 (33%) patients. At presentation, each 3 (50%) patients were diagnosed as stage I and IV, respectively. In initial TETR-PCR according to the original protocol, Cp DNA was detected in none of 6 patients. When TETR-PCR was performed with modification of annealing temperature, Cp DNA was identified in 4 (67%) patients. Direct sequencing could be performed in 3 of 4 positive cases, and specificity of amplified fragments was confirmed.
Summary
PCR optimization with modification of annealing temperature could improve the detection of Cp DNA in patients with OAML. Future prospective validation in a larger sample size is necessary.
Keyword(s): Infection, MALT lymphoma
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