Modified Carba NP test for the detection of carbapenemase production in gram-negative rods: optimized handling of multiple samples

Campana, Eloiza H.; Chuster, Stephanie G.; Silva, Isadora R. da; Paschoal, Raphael P.; Bonelli, Raquel R.; Moreira, Beatriz M.; Picão, Renata C.

doi:10.1016/j.bjm.2016.09.015

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Brazilian Journal of Microbiology

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Medical Microbiology, Short Communication • Braz. J. Microbiol. 48 (2) • Apr-Jun 2017 • https://doi.org/10.1016/j.bjm.2016.09.015 copy

Modified Carba NP test for the detection of carbapenemase production in gram-negative rods: optimized handling of multiple samples

Authorship SCIMAGO INSTITUTIONS RANKINGS

Abstract

The modified Carba NP test presented here may be a valuable tool for laboratories interested in investigating a large number of carbapenemase-producing bacteria in a less-costly way. The test was evaluated against 48 carbapenemase-producing and carbapenemase-non-producing gram-negative bacteria. No false–positive results were obtained, but false-negative results were observed with OXA-23- and GES-carbapenemase-producing isolates. Aeromonas sp. are not testable by Modified Carba NP.

Keywords:
Carbapenemases; Diagnostics; Gram-negative bacteria

Isolates		Species^a a Identification determined by MALDI-TOF MS (Bruker, Germany).	Beta-lactamase (Amber class)^b b β-Lactamases production was assessed by PCR and sequencing assays.	Modified Carba NP test result	Protein concentration (µg/mL ± standard deviation)^c c At the least diluted sample showing positive result.	Highest dilution yielding positive result
Carbapenemase producers	1	Klebsiella pneumoniae	KPC-2 (A)	Positive	<20 ± 5.48	1:128
	2	Kluyvera sp.	KPC-2 (A)	Positive	<20 ± 2.04	1:128
	3	Citrobacter sp.	KPC-2 (A)	Positive	<20 ± 2.31	1:256
	4	Enterobacter cloacae	KPC-2 (A)	Positive	107.39 ± 2.45	1:16
	5	Enterobacter cloacae	KPC-2 (A)	Positive	35.73 ± 21.13	1:64
	6	Klebsiella pneumoniae	KPC-2 (A)	Positive	<20 ± 12.14	1:64
	7	Aeromonas sp.	KPC-2 (A)	Inconclusive^d d Inconclusive: red-to-yellow color change was observed in both imipenem - and + solutions.	-	-
	8	Aeromonas sp.	KPC-like (A)	Inconclusive	-	-
	9	Aeromonas sp.	KPC-like (A)	Inconclusive	-	-
	10	Aeromonas sp.	KPC-like (A)	Inconclusive	-	-
	11	Aeromonas sp.	KPC-like (A)	Inconclusive	-	-
	12	Aeromonas sp.	KPC-like (A)	Inconclusive	-	-
	13	Enterobacter aerogenes	KPC-2 (A)	Positive	24.42 ± 3.8	1:8
	14	Pseudomonas aeruginosa	SPM-1 (B)	Positive	138.09 ± 4.36	1:32
	15	Pseudomonas aeruginosa	SPM-1 (B)	Positive	78.81 ± 4.81	1:64
	16	Pseudomonas aeruginosa	VIM-1 (B)	Positive	397.27 ± 24.29	1:8
	17	Acinetobacter baumannii	SIM-1 (B)	Positive	<20 ± 0.45	1:32
	18	Pseudomonas aeruginosa	GIM-1 (B)	Positive	63.3 ± 2.61	1:128
	19	Klebsiella pneumoniae	NDM-1 (B)	Positive	<20 ± 1.15	1:512
	20	Klebsiella pneumoniae	IMP-1 (B)	Positive	<20 ± 7.92	1:512
	21	Pseudomonas aeruginosa	IMP-18 (B)	Positive	177.15 ± 19.05	1:64
	22	Acinetobacter baumannii	OXA-24 (D)	Positive	570.85 ± 20.56	Undiluted
	23	Klebsiella oxytoca	OXA-48 (D)	Positive	583.22 ± 16.49	Undiluted
	24	Acinetobacter sp.	OXA-58 (D)	Positive	1045.91 ± 76.27	Undiluted
	25	Acinetobacter baumannii	OXA-143 (D)	Positive	452.67 ± 26.04	Undiluted
	26	Acinetobacter baumannii	OXA-23 (D)^e e This isolate show ISAba-1 positioned upstream and in the opposite transcriptional direction of bla OXA-23.	Negative	-	-
	27	Acinetobacter baumannii	OXA-23 (D)^f f This isolate does not show ISAba-1 positioned upstream bla OXA-23.	Negative	-	-
	28	Enterobacter cloacae	GES-5 (A)	Negative	-	-
	29	Enterobacter cloacae	GES-5 (A)	Negative	-	-
	30	Klebsiella peumoniae	GES-16 (A)	Negative	-	-
	31	Enterobacter cloacae	GES-16 (A)	Negative	-	-
	32	Enterobacter cloacae	GES-16 (A)	Negative	-	-
Carbapenemase non-producers	C1	Pseudomonas aeruginosa	GES-1 (A)	Negative	-	-
	C2	Klebsiella pneumoniae	CTX-M-2 (A)	Negative	-	-
	C3	Pseudomonas aeruginosa	OXA-18 (D)	Negative	-	-
	C4	Escherichia coli	DHA-1 (C)	Negative	-	-
	C5	Escherichia coli	CTX-M-15 (A)	Negative	-	-
	C6	Escherichia coli	CTX-M-8 (A)	Negative	-	-
	C7	Escherichia coli	FOX-5 (C)	Negative	-	-
	C8	Escherichia coli	MIR-1 (C)	Negative	-	-
	C9	Escherichia coli	TEM-1 (A)	Negative	-	-
	C10	Aeromonas sp.	GES-like (A)	Inconclusive	-	-
	C11	Aeromonas sp.	GES-like (A)	Inconclusive	-	-
	C12	Aeromonas sp.	None	Inconclusive	-	-
	C13	Aeromonas sp.	None	Inconclusive	-	-
	C14	Aeromonas sp.	None	Inconclusive	-	-
	C15	Pseudomonas aeruginosa ATCC	None	Negative	-	-
	C16	Escherichia coli J53	None	Negative	-	-

Sociedade Brasileira de Microbiologia USP - ICB III - Dep. de Microbiologia, Sociedade Brasileira de Microbiologia, Av. Prof. Lineu Prestes, 2415, Cidade Universitária, 05508-900 São Paulo, SP - Brasil, Ramal USP 7979, Tel. / Fax: (55 11) 3813-9647 ou 3037-7095 - São Paulo - SP - Brazil
E-mail: bjm@sbmicrobiologia.org.br

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[1] *Corresponding author at: Instituto de Microbiologia, Universidade Federal do Rio de Janeiro, Cidade Universitária, Av. Carlos Chagas Filho, 373 – Centro de Ciências da Saúde, Bloco I, Rio de Janeiro, RJ 21941-902, Brazil. E-mail: elocampana@gmail.com (E.H. Campana).