The identification of H. influenzae was based on the requirement of both X and V factor on blood agar. The first three patients were used to assess reproducibility. Nearly 55% of bronchiectasis patients are chronically infected with H. influenzae, which is the most common isolated species from sputum [1-3]. Only patients that cultured H. influenzae alone were included in the study. In this study we only included patients with bronchiectasis confirmed by chest CT according to the features described by Naidich et al. Sputum was collected and each sputum sample was immediately aliquot into separate sterile containers. The aim of the study was to assess the optimum sputum storage conditions for the isolation of Haemophilus influenzae from microbiological culture in patients with bronchiectasis. A paired t-test or Wilcoxon analysis for two groups was performed. The final 6 patient sputum samples were stored in 25% glycerol, 50% skimmed milk and neat at 4°C, -20°C and -70°C for 7 days before processing. 3 specimens from each sample were stored neat at 4°C, -20°C and -70°C (Figure 1C). Of the OMS cultures, 37 (62%) were positive, 22 (36%) were negative, and one (2%) was contaminated. Currently, long-term transport and storage of sputum samples typically requires reliable and continuous access to refrigeration to maintain sample integrity at the level required for smear, culture, and molecular TB diagnostics . Refrigerate and bring the container to the laboratory within one hour of collection. 11 samples were stored short term (2, 4, 6, 24 and 48 hours) at room temperature, 4°C, -20°C and -70°C (Figures 1A and 1B). 98 0 obj << /Linearized 1 /O 100 /H [ 821 708 ] /L 243564 /E 72426 /N 25 /T 241486 >> endobj xref 98 19 0000000016 00000 n ml-1) and viable rates of 91%. Longer term storage of sputum specimens173 samples prior to transport from the field site. They found that 82% of Pseudomonas aeruginosa and 60% of Achromobacter were killed by a single freezing and thawing cycle [9]. ml-1. The viable rates for specimens stored in glycerol at 4°C, -20°C and -70°C was 0.2%, 13.9% and 91%. 1 specimen from each sample was processed immediately after collection, 3 specimens from each sample were stored with 25% glycerol at 4°C, -20°C and -70°C. 0000045945 00000 n Out of the 20 patients, 3 were recruited to assess reproducibility for quantitative microbiology (2 females and 1 male, aged 67, 79 and 35 years). The objectives were to: (1) assess the performance of OMS for transporting sputum from peripheral sites without cold chain stabilization; and (2) compare with Nepal’s standard of care (SOC) for Mycobacterium tuberculosis smear and culture diagnostics. The storage medium, however, can influence H. influenzae viability during freezing and thawing. D (b), the viable rates of samples stored in different conditions. Background: The optimum storage condition is not known for the isolation of H. influenzae in bronchiectasis patient sputum samples, the most common pathogen among bronchiectasis patients. "2Fjr!4Z��M�Ւ�C�QhVAf���g�-��9,E�&�\�P���%s9��a�y$(�Z�>Kͣ�=�Z`��T�8��6Ѡة��+���)����WG�ڭSw��T2��\5�`�����T�Dԫ=�T(]��|A�&;(��E U�?����1�20bH`0�!�÷���+U�+h���Un&K�}��f�0����M��{Ց[F&Dr��ӧ�F�.�\a"�\Dj[���7�3R\��l�Ć���R�Rx�E��ի~_ߺ�p�p���8�������[G�%��8�����fY�]m֪|��'���5ݫ�rV��q_��X��ߺ�+�O��0���{��)����k�U�ڹ�H���l�櫡�>8��欚��V�A����f�]+��]d+x�_�ۤ��i�vr5�$Պl7Zkx̩����Х�;��9��$��]O�հ)���/zk]������U=��ܙS�-��;`�ptt I�������l ����XJJ0&KP��`Z�HFA���.Pa���rDL�I�b�*bR ˀ�"����8*2�]H dyI Ideally, a sputum specimen should be 3–5ml in volume, although smaller quantities are acceptable if … We excluded current or ex-smokers who stopped smoking less than one year ago or ex-smokers with a history of more than 20 pack-years of smoking.
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