Nucleic Acid Amplification Testing and Sequencing Combined with Acid-Fast Staining in Needle Biopsy Lung Tissues for the Diagnosis of Smear-Negative Pulmonary Tuberculosis

<div><p>Background</p><p>Smear-negative pulmonary tuberculosis (PTB) is common and difficult to diagnose. In this study, we investigated the diagnostic value of nucleic acid amplification testing and sequencing combined with acid-fast bacteria (AFB) staining of needle biopsy lung tissues for patients with suspected smear-negative PTB.</p><p>Methods</p><p>Patients with suspected smear-negative PTB who underwent percutaneous transthoracic needle biopsy between May 1, 2012, and June 30, 2015, were enrolled in this retrospective study. Patients with AFB in sputum smears were excluded. All lung biopsy specimens were fixed in formalin, embedded in paraffin, and subjected to acid-fast staining and tuberculous polymerase chain reaction (TB-PCR). For patients with positive AFB and negative TB-PCR results in lung tissues, probe assays and 16S rRNA sequencing were used for identification of nontuberculous mycobacteria (NTM). The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and diagnostic accuracy of PCR and AFB staining were calculated separately and in combination.</p><p>Results</p><p>Among the 220 eligible patients, 133 were diagnosed with TB (men/women: 76/57; age range: 17–80 years, confirmed TB: 9, probable TB: 124). Forty-eight patients who were diagnosed with other specific diseases were assigned as negative controls, and 39 patients with indeterminate final diagnosis were excluded from statistical analysis. The sensitivity, specificity, PPV, NPV, and accuracy of histological AFB (HAFB) for the diagnosis of smear-negative were 61.7% (82/133), 100% (48/48), 100% (82/82), 48.5% (48/181), and 71.8% (130/181), respectively. The sensitivity, specificity, PPV, and NPV of histological PCR were 89.5% (119/133), 95.8% (46/48), 98.3% (119/121), and 76.7% (46/60), respectively, demonstrating that histological PCR had significantly higher accuracy (91.2% [165/181]) than histological acid-fast staining (71.8% [130/181]), <i>P</i> < 0.001. Parallel testing of histological AFB staining and PCR showed the sensitivity, specificity, PPV, NPV, and accuracy to be 94.0% (125/133), 95.8% (46/48), 98.4% (125/127), 85.2% (46/54), and 94.5% (171/181), respectively. Among patients with positive AFB and negative PCR results in lung tissue specimens, two were diagnosed with NTM infections (<i>Mycobacterium avium-intracellulare</i> complex and <i>Mycobacterium kansasii</i>).</p><p>Conclusion</p><p>Nucleic acid amplification testing combined with acid-fast staining in lung biopsy tissues can lead to early and accurate diagnosis in patients with smear-negative pulmonary tuberculosis. For patients with positive histological AFB and negative tuberculous PCR results in lung tissue, NTM infection should be suspected and could be identified by specific probe assays or 16S rRNA sequencing.</p></div

Clinical characteristics of the 10 HAFB-positive and TB-PCR-negative patients.

<p>Clinical characteristics of the 10 HAFB-positive and TB-PCR-negative patients.</p

Baseline characteristics of the study patients.

<p>Baseline characteristics of the study patients.</p

Diagnostic performance of HAFB and real-time PCR for analysis of lung biopsy specimens.

<p>Diagnostic performance of HAFB and real-time PCR for analysis of lung biopsy specimens.</p

Air Cushion Convection Inhibiting Icing of Self-Cleaning Surfaces

Anti-icing surfaces/interfaces are of considerable importance in various engineering fields under natural freezing environment. Although superhydrophobic self-cleaning surfaces show good anti-icing potentials, promotion of these surfaces in engineering applications seems to enter a “bottleneck” stage. One of the key issues is the intrinsic relationship between superhydrophobicity and icephobicity is unclear, and the dynamic action mechanism of “air cushion” (a key internal factor for superhydrophobicity) on icing suppression was largely ignored. Here we report that icing inhibition (i.e., icing-delay) of self-cleaning surfaces is mainly ascribed to air cushion and its convection. We experimentally found air cushion on the porous self-cleaning coating under vacuum environments and on the water/ice-coating interface at low temperatures. The icing-delay performances of porous self-cleaning surfaces compared with bare substrate, up to 10–40 min under 0 to ∼−4 °C environments close to freezing rain, have been accurately real-time recorded by a novel synergy method including high-speed photography and strain sensing voltage. Based on the experimental results, we innovatively propose a physical model of “air cushion convection inhibiting icing”, which envisages both the static action of trapped air pocket without air flow and dynamic action of air cushion convection. Gibbs free energy of water droplets increased with the entropy of air derived from heat and mass transfer between warmer air underneath water droplets and colder surrounding air, resulting in remarkable ice nucleation delay. Only when air cushion convection disappears can ice nucleation be triggered on suitable Gibbs free energy conditions. The fundamental understanding of air cushion on anti-icing is an important step toward designing optimal anti-icing surfaces for practical engineering application