Detection of Nocardia, Streptomyces and Rhodococcus from bronchoalveolar lavage specimens of patients with HIV by Multiplex PCR Assay

Abstract

Background: Nocardia, Streptomyces and Rhodococcus are life threatening opportunistic pathogens under immunodeficiency conditions, particularly among patients infected with HIV. Rapid and accurate detection of these infections can improve immune health quality, patient management and appropriate treatment. The aim of this study was to design a novel multiplex-PCR assay for rapid diagnosis of these three organisms directly from bronchoalveolar lavage (BAL) specimens of patients infected with HIV.Methods: The genus specific primers were designed for directdetection of Nocardia, Streptomyces and Rhodococcus in a single tube multiplex PCR. This PCR specifically amplified the target genes from pure cultures. It subsequently was applied on BAL specimens of 29 HIV positive patients that had previously been culture negative for actinomycete bacteria, of which Nocardia, Streptomyces and Rhodococcus are members.Results: Of 29 respiratory clinical specimens, there were positive for Nocardia spp. and one was positive for Streptomyces spp using the multiplex PCR assay. The sequencing of the PCR products identified the species as Nocardia cyriacigeorgica (n=2), Nocardia farcinica and Streptomyces albus.Conclusion: This novel multiplex PCR assay yielded reliable results for accurate identification of Nocardia, Streptomyces and Rhodococcus from BAL while the results of bacterial culture were negative.

Two complete 1918 influenza A/H1N1 pandemic virus genomes characterized by next-generation sequencing using RNA isolated from formalin-fixed, paraffin-embedded autopsy lung tissue samples along with evidence of secondary bacterial co-infection

“…The genus Rhodococcus , together with the genus Nocardia , is a member of family Nocardiaceae and is classified in the suborder Corynebacterineae ( 54 ). Pulmonary nocardiosis is an invasive infection caused by Nocardia , a partial acid-fast microorganism, and the common route of pulmonary infection is inhalation of contaminated aerosols from environmental sources ( 55 ). Rhodococcus is a genus of aerobic, nonsporulating, nonmotile Gram-positive bacteria, mostly found in a broad range of environments, including soils, groundwater, marine sediments, animal dung, and the guts of insects as well as from healthy and diseased animals and plants ( 56 , 57 ).…”

Evaluation of the Diagnostic Accuracy and Clinical Utility of Fungal Profile Plus Polymerase Chain Reaction Assay in Pulmonary Infections

“…One study on BALF PCR for Mucorales combined PCR and high-resolution melt analysis for a combined 90% sensitivity [ 23 ]. One small study of Nocardia PCR reported 100% sensitivity by multiplex PCR in patients with HIV (n = 4), while another study in lung transplant patients detected Nocardia in 5 of 29 (17.2%) BALF specimens; however, none were associated with clinical disease [ 24 , 25 ].…”

Genotypic and phenotypic prevalence of Nocardia species in Iran: First systematic review and meta-analysis of data accumulated over years 1992-2021

“…Then based on the abstract evaluation, 55 articles were excluded (3 review articles, 27 case reports, 8 letters to the editor, and 17 were related to non-clinical Nocardia specimens). So, according to quality assessment criteria and inclusion/exclusion criteria data, a remaining 20 most-related articles were included in present study [10][11][12][13][14][15][16][17][18][19][20][21][22][23][24][25][26][27][28][29]. Among 20 articles involving a total of 338 Nocardia isolates, the prevalence of Nocardia spp.…”