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Isolation of bacterial DNA followed by sequencing and differing cytokine response in peritoneal dialysis effluent help in identifying bacteria in culture negative peritonitis
Author(s) -
Prasad Narayan,
Singh Kamini,
Gupta Amit,
Prasad Kashi Nath
Publication year - 2018
Publication title -
nephrology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.752
H-Index - 61
eISSN - 1440-1797
pISSN - 1320-5358
DOI - 10.1111/nep.12969
Subject(s) - peritonitis , bacteria , microbiology and biotechnology , gram negative bacteria , peritoneal dialysis , gram positive bacteria , polymerase chain reaction , antibiotics , microbiological culture , medicine , biology , escherichia coli , gene , genetics
Aim The treatment of peritoneal dialysis related culture negative peritonitis is empirical which increases the cost of therapy and moreover antibiotic resistance. We aimed the study to isolate bacterial DNA from PD effluent and indentify bacteria causing peritonitis in culture negative situations. We have also studied the cytokine response with different bacteria causing peritonitis. Methods We have isolated bacterial DNA from PD effluent of culture negative and culture positive peritonitis patients. Bacterial DNA was subjected to polymerase chain reaction using universal bacteria specific primers and subsequently to Gram type specific primers for the differentiation of the etiologic agents into Gram‐positive and Gram‐negative. The amplified products were sequenced and subjected to blast search to identify agent at genus/ species level. Results Of the 30 molecular method positive samples, 16 (53.33%) samples were positive for Gram‐negative bacteria and 4 (13.33%) for Gram‐positive, while the remaining10 (33.33%) were positive for both Gram‐positive and Gram‐negative bacteria. We have found organisms that usually do not grow on normal culture methods. TNF‐α was significantly associated with Gram‐positive peritonitis and regulatory cytokine IL‐10 with Gram‐negative peritonitis. Conclusions The molecular techniques are helpful in detecting and identifying organisms from culture negative PD effluent.